Scholars@Duke
Arno Lee Greenleaf

Arno Lee Greenleaf

Professor Emeritus of Biochemistry
Biochemistry
arno@duke.edu
Duke Box 3711, Durham, NC 27710
0034 CARL Building, Duke Box 3711, Durham, NC 27710

Selected Publications

CDK12 Activity-Dependent Phosphorylation Events in Human Cells.

Journal Article Biomolecules · October 22, 2019 We asked whether the C-terminal repeat domain (CTD) kinase, CDK12/CyclinK, phosphorylates substrates in addition to the CTD of RPB1, using our CDK12analog-sensitive HeLa cell line to investigate CDK12 activity-dependent phosphorylation events in human cell ... Full text Link to item Cite

Human CDK12 and CDK13, multi-tasking CTD kinases for the new millenium.

Journal Article Transcription · April 2019 As the new millennium began, CDK12 and CDK13 were discovered as nucleotide sequences that encode protein kinases related to cell cycle CDKs. By the end of the first decade both proteins had been qualified as CTD kinases, and it was emerging that both are h ... Full text Link to item Cite

Covalent targeting of remote cysteine residues to develop CDK12 and CDK13 inhibitors.

Journal Article Nat Chem Biol · October 2016 Cyclin-dependent kinases 12 and 13 (CDK12 and CDK13) play critical roles in the regulation of gene transcription. However, the absence of CDK12 and CDK13 inhibitors has hindered the ability to investigate the consequences of their inhibition in healthy cel ... Full text Link to item Cite

Engineering an analog-sensitive CDK12 cell line using CRISPR/Cas.

Journal Article Biochim Biophys Acta · September 2015 The RNA Polymerase II C-terminal domain (CTD) kinase CDK12 has been implicated as a tumor suppressor and regulator of DNA damage response genes. Although much has been learned about CDK12 and its activity, due to the lack of a specific inhibitor and the co ... Full text Link to item Cite

Expression, purification, and identification of associated proteins of the full-length hCDK12/CyclinK complex.

Journal Article J Biol Chem · January 16, 2015 The coupling of transcription and associated processes has been shown to be dependent on the RNA polymerase II (RNAPII) C-terminal repeat domain (CTD) and the phosphorylation of the heptad repeats of which it is composed (consensus sequence Y1S2P3T4S5P6S7) ... Full text Link to item Cite

Specific interaction of the transcription elongation regulator TCERG1 with RNA polymerase II requires simultaneous phosphorylation at Ser2, Ser5, and Ser7 within the carboxyl-terminal domain repeat.

Journal Article J Biol Chem · April 12, 2013 The human transcription elongation regulator TCERG1 physically couples transcription elongation and splicing events by interacting with splicing factors through its N-terminal WW domains and the hyperphosphorylated C-terminal domain (CTD) of RNA polymerase ... Full text Link to item Cite

A DNA damage response system associated with the phosphoCTD of elongating RNA polymerase II.

Journal Article PLoS One · 2013 RNA polymerase II translocates across much of the genome and since it can be blocked by many kinds of DNA lesions, detects DNA damage proficiently; it thereby contributes to DNA repair and to normal levels of DNA damage resistance. However, the components ... Full text Link to item Cite

Proteomic analysis of mitotic RNA polymerase II reveals novel interactors and association with proteins dysfunctional in disease.

Journal Article Mol Cell Proteomics · June 2012 RNA polymerase II (RNAPII) transcribes protein-coding genes in eukaryotes and interacts with factors involved in chromatin remodeling, transcriptional activation, elongation, and RNA processing. Here, we present the isolation of native RNAPII complexes usi ... Full text Link to item Cite

Cotranscriptional association of mRNA export factor Yra1 with C-terminal domain of RNA polymerase II.

Journal Article J Biol Chem · October 21, 2011 The unique C-terminal domain (CTD) of RNA polymerase II, composed of tandem heptad repeats of the consensus sequence YSPTSPS, is subject to differential phosphorylation throughout the transcription cycle. Several RNA processing factors have been shown to b ... Full text Link to item Cite

Phosphorylation of RNAPII: To P-TEFb or not to P-TEFb?

Journal Article Transcription · May 2011 The C-terminal domain of RNA polymerase II undergoes a cycle of phosphorylation which allows it to temporally couple transcription with transcription-associated processes. The characterization of hitherto unrecognized metazoan elongation phase CTD kinase a ... Full text Link to item Cite

cis-Proline-mediated Ser(P)5 dephosphorylation by the RNA polymerase II C-terminal domain phosphatase Ssu72.

Journal Article J Biol Chem · February 18, 2011 RNA polymerase II coordinates co-transcriptional events by recruiting distinct sets of nuclear factors to specific stages of transcription via changes of phosphorylation patterns along its C-terminal domain (CTD). Although it has become increasingly clear ... Full text Link to item Cite

Updating the CTD Story: From Tail to Epic.

Journal Article Genet Res Int · 2011 Eukaryotic RNA polymerase II (RNAPII) not only synthesizes mRNA but also coordinates transcription-related processes via its unique C-terminal repeat domain (CTD). The CTD is an RNAPII-specific protein segment consisting of repeating heptads with the conse ... Full text Link to item Cite

RECQ5 helicase associates with the C-terminal repeat domain of RNA polymerase II during productive elongation phase of transcription.

Journal Article Nucleic Acids Res · December 2010 It is known that transcription can induce DNA recombination, thus compromising genomic stability. RECQ5 DNA helicase promotes genomic stability by regulating homologous recombination. Recent studies have shown that RECQ5 forms a stable complex with RNA pol ... Full text Link to item Cite

Genetic organization, length conservation, and evolution of RNA polymerase II carboxyl-terminal domain.

Journal Article Mol Biol Evol · November 2010 With a simple tandem iterated sequence, the carboxyl terminal domain (CTD) of eukaryotic RNA polymerase II (RNAP II) serves as the central coordinator of mRNA synthesis by harmonizing a diversity of sequential interactions with transcription and processing ... Full text Link to item Cite

CDK12 is a transcription elongation-associated CTD kinase, the metazoan ortholog of yeast Ctk1.

Journal Article Genes Dev · October 15, 2010 Drosophila contains one (dCDK12) and humans contain two (hCDK12 and hCDK13) proteins that are the closest evolutionary relatives of yeast Ctk1, the catalytic subunit of the major elongation-phase C-terminal repeat domain (CTD) kinase in Saccharomyces cerev ... Full text Link to item Cite

The phosphoCTD-interacting domain of Topoisomerase I.

Journal Article Biochem Biophys Res Commun · June 18, 2010 The N-terminal domain (NTD) of Drosophila melanogaster (Dm) Topoisomerase I has been shown to bind to RNA polymerase II, but the domain of RNAPII with which it interacts is not known. Using bacterially-expressed fusion proteins carrying all or half of the ... Full text Link to item Cite

Comparative genome-wide screening identifies a conserved doxorubicin repair network that is diploid specific in Saccharomyces cerevisiae.

Journal Article PLoS One · June 8, 2009 The chemotherapeutic doxorubicin (DOX) induces DNA double-strand break (DSB) damage. In order to identify conserved genes that mediate DOX resistance, we screened the Saccharomyces cerevisiae diploid deletion collection and identified 376 deletion strains ... Full text Link to item Cite

The essential sequence elements required for RNAP II carboxyl-terminal domain function in yeast and their evolutionary conservation.

Journal Article Mol Biol Evol · April 2008 The carboxyl-terminal domain (CTD) of eukaryotic RNA polymerase II is the staging platform for numerous proteins involved in transcription initiation, mRNA processing, and general coordination of nuclear events. Concordant with these central roles in cellu ... Full text Link to item Cite

Yeast screens identify the RNA polymerase II CTD and SPT5 as relevant targets of BRCA1 interaction.

Journal Article PLoS One · January 16, 2008 BRCA1 has been implicated in numerous DNA repair pathways that maintain genome integrity, however the function responsible for its tumor suppressor activity in breast cancer remains obscure. To identify the most highly conserved of the many BRCA1 functions ... Full text Open Access Link to item Cite

Phosphorylation and functions of the RNA polymerase II CTD.

Journal Article Genes Dev · November 1, 2006 The C-terminal repeat domain (CTD), an unusual extension appended to the C terminus of the largest subunit of RNA polymerase II, serves as a flexible binding scaffold for numerous nuclear factors; which factors bind is determined by the phosphorylation pat ... Full text Link to item Cite

Solution structure of the Set2-Rpb1 interacting domain of human Set2 and its interaction with the hyperphosphorylated C-terminal domain of Rpb1.

Journal Article Proc Natl Acad Sci U S A · December 6, 2005 The phosphorylation state of the C-terminal repeat domain (CTD) of the largest subunit of RNA polymerase II changes as polymerase transcribes a gene, and the distinct forms of the phospho-CTD (PCTD) recruit different nuclear factors to elongating polymeras ... Full text Link to item Cite

A novel domain in Set2 mediates RNA polymerase II interaction and couples histone H3 K36 methylation with transcript elongation.

Journal Article Mol Cell Biol · April 2005 Histone methylation and the enzymes that mediate it are important regulators of chromatin structure and gene transcription. In particular, the histone H3 lysine 36 (K36) methyltransferase Set2 has recently been shown to associate with the phosphorylated C- ... Full text Link to item Cite

Expanding the functional repertoire of CTD kinase I and RNA polymerase II: novel phosphoCTD-associating proteins in the yeast proteome.

Journal Article Biochemistry · December 21, 2004 CTD kinase I (CTDK-I) of Saccharomyces cerevisiae is required for normal phosphorylation of the C-terminal repeat domain (CTD) on elongating RNA polymerase II. To elucidate cellular roles played by this kinase and the hyperphosphorylated CTD (phosphoCTD) i ... Full text Link to item Cite

C-terminal repeat domain kinase I phosphorylates Ser2 and Ser5 of RNA polymerase II C-terminal domain repeats.

Journal Article J Biol Chem · June 11, 2004 The C-terminal repeat domain (CTD) of the largest subunit of RNA polymerase II is composed of tandem heptad repeats with consensus sequence Tyr1-Ser2-Pro3-Thr4-Ser5-Pro6-Ser7. In yeast, this heptad sequence is repeated about 26 times, and it becomes hyperp ... Full text Link to item Cite

Identifying phosphoCTD-associating proteins.

Journal Article Methods Mol Biol · 2004 The C-terminal repeat domain (CTD) of the largest subunit of RNA polymerase II is hyperphosphorylated during transcription elongation. The phosphoCTD is known to bind to a subset of RNA processing factors and to several other nuclear proteins, thereby posi ... Full text Link to item Cite

Getting a grip on the CTD of Pol II.

Journal Article Structure · August 2003 The first structure of a pre-mRNA processing factor bound to heptad repeats from the C-terminal domain of RNA polymerase II is revealed in a crystal of capping guanylyltransferase complexed with a four-repeat phosphopeptide. ... Full text Link to item Cite

The RNA polymerase II CTD kinase CTDK-I affects pre-mRNA 3' cleavage/polyadenylation through the processing component Pti1p.

Journal Article Mol Cell · December 2002 Featured Publication There are several kinases in Saccharomyces cerevisiae that phosphorylate the CTD of RNA polymerase II, but specific and distinct functions of the phospho-CTDs generated by the different kinases are not well understood. A genetic screen for suppressors of l ... Full text Link to item Cite

Hyperphosphorylated C-terminal repeat domain-associating proteins in the nuclear proteome link transcription to DNA/chromatin modification and RNA processing.

Journal Article Mol Cell Proteomics · August 2002 Featured Publication Using an interaction blot approach to search in the human nuclear proteome, we identified eight novel proteins that bind the hyperphosphorylated C-terminal repeat domain (phosphoCTD) of RNA polymerase II. Unexpectedly, five of the new phosphoCTD-associatin ... Full text Link to item Cite

Co-transcriptional splicing of pre-messenger RNAs: considerations for the mechanism of alternative splicing.

Journal Article Gene · October 17, 2001 Featured Publication Nascent transcripts are the true substrates for many splicing events in mammalian cells. In this review we discuss transcription, splicing, and alternative splicing in the context of co-transcriptional processing of pre-mRNA. The realization that splicing ... Full text Link to item Cite

Phosphorylation of RNA polymerase II CTD fragments results in tight binding to the WW domain from the yeast prolyl isomerase Ess1.

Journal Article Biochemistry · July 24, 2001 Featured Publication The yeast prolyl isomerase, Ess1, has recently been shown to interact via its WW domain with the hyperphosphorylated form of the RNA polymerase II C-terminal domain (CTD). We have investigated folding of the Ess1 WW domain and its binding to peptides repre ... Full text Link to item Cite

Juglone, an inhibitor of the peptidyl-prolyl isomerase Pin1, also directly blocks transcription.

Journal Article Nucleic Acids Res · February 1, 2001 The C-terminal domain (CTD) of the large subunit of RNA polymerase II plays a role in transcription and RNA processing. Yeast ESS1, a peptidyl-prolyl cis/trans isomerase, is involved in RNA processing and can associate with the CTD. Using several types of ... Full text Link to item Cite

The splicing factor, Prp40, binds the phosphorylated carboxyl-terminal domain of RNA polymerase II.

Journal Article J Biol Chem · December 22, 2000 Featured Publication We showed previously that the WW domain of the prolyl isomerase, Ess1, can bind the phosphorylated carboxyl-terminal domain (phospho-CTD) of the largest subunit of RNA Polymerase II. Analysis of phospho-CTD binding by four other WW domain-containing Saccha ... Full text Link to item Cite

Protein-interaction modules that organize nuclear function: FF domains of CA150 bind the phosphoCTD of RNA polymerase II.

Journal Article Proc Natl Acad Sci U S A · August 1, 2000 Featured Publication An approach for purifying nuclear proteins that bind directly to the hyperphosphorylated C-terminal repeat domain (CTD) of RNA polymerase II was developed and used to identify one human phosphoCTD-associating protein as CA150. CA150 is a nuclear factor imp ... Full text Link to item Cite

Kin28, the TFIIH-associated carboxy-terminal domain kinase, facilitates the recruitment of mRNA processing machinery to RNA polymerase II.

Journal Article Mol Cell Biol · January 2000 The cotranscriptional placement of the 7-methylguanosine cap on pre-mRNA is mediated by recruitment of capping enzyme to the phosphorylated carboxy-terminal domain (CTD) of RNA polymerase II. Immunoblotting suggests that the capping enzyme guanylyltransfer ... Full text Link to item Cite

Phospho-carboxyl-terminal domain binding and the role of a prolyl isomerase in pre-mRNA 3'-End formation.

Journal Article J Biol Chem · October 29, 1999 Featured Publication A phospho-carboxyl-terminal domain (CTD) affinity column created with yeast CTD kinase I and the CTD of RNA polymerase II was used to identify Ess1/Pin1 as a phospho-CTD-binding protein. Ess1/Pin1 is a peptidyl prolyl isomerase involved in both mitotic reg ... Full text Link to item Cite

Heat-shock-specific phosphorylation and transcriptional activity of RNA polymerase II.

Journal Article Exp Cell Res · July 10, 1998 The carboxyl-terminal domain (CTD) of the largest RNA polymerase II (pol II) subunit is a target for extensive phosphorylation in vivo. Using in vitro kinase assays it was found that several different protein kinases can phosphorylate the CTD including the ... Full text Link to item Cite

Protonation of the neutral repeats of the RNA polymerase II CTD.

Journal Article Biochem Biophys Res Commun · April 7, 1998 The CTD (carboxy-terminal repeat domain) of the largest subunit of RNA Polymerase II in most eukaryotes consists of from 26 to 52 seven amino acid repeats, the consensus sequence of which is YSPTSPS. Even though this consensus repeat does not contain resid ... Full text Link to item Cite

Assaying CTD kinases in vitro and phosphorylation-modulated properties of RNA polymerase II in vivo.

Journal Article Methods · July 1997 The functional properties of RNA polymerase II are modulated by hyperphosphorylation of its unique C-terminal repeat domain (CTD). A number of enzymes with CTD kinase activity have been identified, and correlations between CTD phosphorylation and RNA polym ... Full text Link to item Cite

Modulation of RNA polymerase II elongation efficiency by C-terminal heptapeptide repeat domain kinase I.

Journal Article J Biol Chem · April 25, 1997 Featured Publication Hyperphosphorylation of the C-terminal heptapeptide repeat domain (CTD) of the RNA polymerase II largest subunit has been suggested to play a key role in regulating transcription initiation and elongation. To facilitate investigating functional consequence ... Full text Link to item Cite

Analyses of promoter-proximal pausing by RNA polymerase II on the hsp70 heat shock gene promoter in a Drosophila nuclear extract.

Journal Article Mol Cell Biol · October 1996 Analyses of Drosophila cells have revealed that RNA polymerase II is paused in a region 20 to 40 nucleotides downstream from the transcription start site of the hsp70 heat shock gene when the gene is not transcriptionally active. We have developed a cell-f ... Full text Link to item Cite

Drosophila RNA polymerase II mutants that affect transcription elongation.

Journal Article J Biol Chem · March 15, 1996 We have examined the properties of two Drosophila RNA polymerase II mutants, C4 and S1, during elongation, pyrophosphorolysis, and DmS-II-stimulated transcript cleavage. The C4 and S1 mutants contain a single amino acid substitution in the largest and seco ... Link to item Cite

Phosphorylation dependence of the initiation of productive transcription of Balbiani ring 2 genes in living cells.

Journal Article Chromosoma · March 1996 Using polytene chromosomes of salivary gland cells of Chironomus tentans, phosphorylation state-sensitive antibodies and the transcription and protein kinase inhibitor 5,6-dichloro-1-beta-D-ribofuranosylbenzimidazole (DRB), we have visualized the chromosom ... Full text Link to item Cite

The yeast carboxyl-terminal repeat domain kinase CTDK-I is a divergent cyclin-cyclin-dependent kinase complex.

Journal Article Mol Cell Biol · October 1995 Featured Publication Saccharomyces cerevisiae CTDK-I is a protein kinase complex that specifically and efficiently hyperphosphorylates the carboxyl-terminal repeat domain (CTD) of RNA polymerase II and is composed of three subunits of 58, 38, and 32 kDa. The kinase is essentia ... Full text Link to item Cite

Functional studies of the carboxy-terminal repeat domain of Drosophila RNA polymerase II in vivo.

Journal Article Genetics · June 1995 To understand the in vivo function of the unique and conserved carboxy-terminal repeat domain (CTD) of RNA polymerase II largest subunit (RpII215), we have studied RNA polymerase II biosynthesis, activity and genetic function in Drosophila RpII215 mutants ... Full text Link to item Cite

Identifying a transcription factor interaction site on RNA polymerase II.

Journal Article Gene Expr · 1995 We have generated a series of fusion proteins carrying portions of subunit IIc, the second largest subunit of Drosophila RNA polymerase I, and have used them in a domain interference assay to identify a fragment of the IIc subunit that carries the binding ... Link to item Cite

Phosphorylation of RNA polymerase II C-terminal domain and transcriptional elongation.

Journal Article Nature · July 7, 1994 The carboxy-terminal domain (CTD) of the large subunit of RNA polymerase II is essential in vivo, and is found in either an unphosphorylated (IIa) or hyperphosphorylated (IIo) form. The Drosophila uninduced hsp70 and hsp26 genes, and the constitutively exp ... Full text Link to item Cite

RNA-POLYMERASE-II CTD PHOSPHORYLATION IN-VIVO

Journal Article JOURNAL OF CELLULAR BIOCHEMISTRY · February 13, 1994 Link to item Cite

Locus-specific variation in phosphorylation state of RNA polymerase II in vivo: correlations with gene activity and transcript processing.

Journal Article Genes Dev · December 1993 Featured Publication To investigate functional differences between RNA polymerases IIA and IIO (Pol IIA and Pol IIO), with hypo- and hyperphosphorylated carboxy-terminal repeat domains (CTDs), respectively, we have visualized the in vivo distributions of the differentially pho ... Full text Link to item Cite

A positive addition to a negative tail's tale.

Journal Article Proc Natl Acad Sci U S A · December 1, 1993 Full text Link to item Cite

Mapping mutations in genes encoding the two large subunits of Drosophila RNA polymerase II defines domains essential for basic transcription functions and for proper expression of developmental genes.

Journal Article Mol Cell Biol · July 1993 We have mapped a number of mutations at the DNA sequence level in genes encoding the largest (RpII215) and second-largest (RpII140) subunits of Drosophila melanogaster RNA polymerase II. Using polymerase chain reaction (PCR) amplification and single-strand ... Full text Link to item Cite

Reverse genetics of Drosophila RNA polymerase II: identification and characterization of RpII140, the genomic locus for the second-largest subunit.

Journal Article Genetics · June 1993 We have used a reverse genetics approach to isolate genes encoding two subunits of Drosophila melanogaster RNA polymerase II. RpII18 encodes the 18-kDa subunit and maps cytogenetically to polytene band region 83A. RpII140 encodes the 140-kDa subunit and ma ... Full text Link to item Cite

Positive patches and negative noodles: linking RNA processing to transcription?

Journal Article Trends Biochem Sci · April 1993 Featured Publication A speculative model is presented that proposes specific mechanisms for effecting co-transcriptional splice site selection in pre-mRNAs. The model envisions that certain splicing factors containing arginine-rich, positively charged regions bind via these po ... Full text Link to item Cite

CTD kinase large subunit is encoded by CTK1, a gene required for normal growth of Saccharomyces cerevisiae.

Journal Article Gene Expr · May 1991 Featured Publication We previously purified a yeast protein kinase that specifically hyperphosphorylates the carboxyl-terminal repeat domain (CTD) of RNA polymerase II largest subunit and showed that this CTD kinase consists of three subunits of 58, 38, and 32 kDa. We have now ... Link to item Cite

The carboxyl-terminal repeat domain of RNA polymerase II is not required for transcription factor Sp1 to function in vitro.

Journal Article J Biol Chem · May 25, 1990 We show that the mammalian transcription Sp1 stimulates accurate transcription in a partially fractionated RNA polymerase II-dependent system from Drosophila cultured cells. Moreover, the extent of stimulation is equal for intact RNA polymerase II (polymer ... Link to item Cite

Properties of a Drosophila RNA polymerase II elongation factor.

Journal Article J Biol Chem · May 25, 1989 We have purified from nuclear extracts of Drosophila Kc cells a 36-kDa protein, DmS-II, which has an effect on the elongation properties of RNA polymerase II. DmS-II stimulates RNA polymerase II during the transcription of double-stranded DNA templates whe ... Link to item Cite

A protein kinase that phosphorylates the C-terminal repeat domain of the largest subunit of RNA polymerase II.

Journal Article Proc Natl Acad Sci U S A · May 1989 The unique C-terminal repeat domain (CTD) of the largest subunit (IIa) of eukaryotic RNA polymerase II consists of multiple repeats of the heptapeptide consensus sequence Tyr-Ser-Pro-Thr-Ser-Pro-Ser. The number of repeats ranges from 26 in yeast to 42 in D ... Full text Link to item Cite

Dynamic interaction between a Drosophila transcription factor and RNA polymerase II.

Journal Article Mol Cell Biol · April 1989 We have purified factor 5, a Drosophila RNA polymerase II transcription factor. Factor 5 was found to be required for accurate initiation of transcription from specific promoters and also had a dramatic effect on the elongation properties of RNA polymerase ... Full text Link to item Cite

Heat shock-induced appearance of RNA polymerase II in karyoskeletal protein-enriched (nuclear "matrix") fractions correlates with transcriptional shutdown in Drosophila melanogaster.

Journal Article J Biol Chem · February 25, 1989 Heat shock in vivo or brief incubation at moderately elevated temperatures (15 min at 37 degrees C) in vitro is required for the structural stability of proteinaceous karyoskeletal elements purified from Drosophila melanogaster (McConnell, M., Whalen, A. M ... Link to item Cite

Analysis of the gene encoding the largest subunit of RNA polymerase II in Drosophila.

Journal Article Mol Gen Genet · January 1989 We have characterized RpII215, the gene encoding the largest subunit of RNA polymerase II in Drosophila melanogaster. DNA sequencing and nuclease S1 analyses provided the primary structure of this gene, its 7 kb RNA and 215 kDa protein products. The amino- ... Full text Link to item Cite

Elongation by Drosophila RNA polymerase II. Transcription of 3'-extended DNA templates.

Journal Article J Biol Chem · July 15, 1988 RNA polymerase II will efficiently initiate transcription on linear duplex DNA which has been extended at its 3' ends by the addition of short stretches of polydeoxycytidine (Kadesch, T. R., and Chamberlin, M. J. (1982) J. Biol. Chem. 257, 5286-5295). We h ... Link to item Cite

The C-terminal repeat domain of RNA polymerase II largest subunit is essential in vivo but is not required for accurate transcription initiation in vitro.

Journal Article Proc Natl Acad Sci U S A · June 1988 DNA sequence analysis of RpII215, the gene that encodes the Mr215,000 subunit of RNA polymerase II (EC 2.7.7.6) in Drosophila melanogaster, reveals that the 3'-terminal exon includes a region encoding a C-terminal domain composed of 42 repeats of a seven-r ... Full text Link to item Cite

Fractionation of transcription factors for RNA polymerase II from Drosophila Kc cell nuclear extracts.

Journal Article J Biol Chem · March 5, 1987 Drosophila Kc cells were utilized to prepare nuclear extracts in which promoter-containing DNA templates were efficiently transcribed by RNA polymerase II. A combination of fractionation schemes was used to identify and partially purify seven activities (f ... Link to item Cite

An activity necessary for in vitro transcription is a DNase inhibitor.

Journal Article Biochimie · 1987 A phosphocellulose flowthrough fraction required for accurate transcription in vitro by RNA polymerase II was found to contain a DNase inhibitor which was necessary to maintain template integrity (Price D.H., Sluder A.E. & Greenleaf A.L. (1987) J. Biol. Ch ... Full text Link to item Cite

Sites of P element insertion and structures of P element deletions in the 5' region of Drosophila melanogaster RpII215.

Journal Article Mol Cell Biol · October 1986 Several P element insertion and deletion mutations near the 5' end of Drosophila melanogaster RpII215 have been examined by nucleotide sequencing. Two different sites of P element insertion, approximately 90 nucleotides apart, have been detected in this re ... Full text Link to item Cite

Isolation of the nuclear gene encoding a subunit of the yeast mitochondrial RNA polymerase.

Journal Article J Biol Chem · August 5, 1986 Antisera directed against the purified 145,000-dalton subunit of the Saccharomyces cerevisiae mitochondrial RNA polymerase have been used to immuno-screen a library of yeast genomic inserts constructed in the fusion protein expression vector, lambda gt11. ... Link to item Cite

Yeast RPO41 gene product is required for transcription and maintenance of the mitochondrial genome.

Journal Article Proc Natl Acad Sci U S A · May 1986 A 4-kilobase DNA fragment carried by a recombinant lambda gt11 bacteriophage appears to contain most of the coding information for the 145-kDa subunit of the Saccharomyces cerevisiae mitochondrial RNA polymerase. The RPO41 gene is located on chromosome VI, ... Full text Link to item Cite

Dye-ligand affinity chromatography of RNA polymerase II.

Journal Article Acta Biochim Biophys Hung · 1986 The binding of wheat germ RNA polymerase II to five different dye-ligand chromatography gels (Matrex gels, Amicon Corp.) was tested. A quantitative binding of the enzyme to four of the gels, namely Dyematrex gels Blue A, Blue B, Red A and Green A was obser ... Link to item Cite

A mutation in the largest subunit of RNA polymerase II alters RNA chain elongation in vitro.

Journal Article J Biol Chem · October 25, 1985 An in vitro transcription system which utilized a semisynthetic DNA template (Kadesch, T. R., and Chamberlin, M. J. (1982) J. Biol. Chem. 257, 5286-5295) was developed and used to compare RNA chain elongation by wild type and mutant RNA polymerases II of D ... Link to item Cite

Structure of the eukaryotic transcription apparatus: features of the gene for the largest subunit of Drosophila RNA polymerase II.

Journal Article Cell · September 1985 The Drosophila melanogaster RpII215 locus encodes the largest subunit of RNA polymerase II. We have now mapped the 7 kb transcript of the locus and have determined that it contains four exons and three introns. By sequencing 2582 nucleotides from the promo ... Full text Link to item Cite

Frequent Imprecise Excision among Reversions of a P Element-Caused Lethal Mutation in Drosophila.

Journal Article Genetics · June 1984 RpII215(D) (50) (= D50) is a lethal mutation caused by the insertion of a 1.3-kb P element 5' to sequences encoding the largest (215 kilodaltons) subunit of Drosophila RNA polymerase II. In dysgenic males D50 reverted to nonlethality at frequencies ranging ... Full text Link to item Cite

Identification, molecular cloning, and mutagenesis of Saccharomyces cerevisiae RNA polymerase genes.

Journal Article Proc Natl Acad Sci U S A · April 1984 Three different regions of Saccharomyces cerevisiae DNA were identified by using as hybridization probe a fragment of Drosophila melanogaster DNA that encodes an RNA polymerase II (EC 2.7.7.6) polypeptide. Two of these regions have been molecularly cloned. ... Full text Link to item Cite

Affinity-purified antibody as a probe of RNA polymerase II subunit structure.

Journal Article J Mol Appl Genet · 1984 Antibodies reactive against mammalian RNA polymerase II were isolated by affinity chromatography. Serum from a goat immunized with Drosophila RNA polymerase II was passed over a column containing covalently coupled calf thymus RNA polymerase II, and reacti ... Link to item Cite

Amanitin-resistant RNA polymerase II mutations are in the enzyme's largest subunit.

Journal Article J Biol Chem · November 25, 1983 A fragment of the Drosophila melanogaster RpIIC4 locus, which encodes the RNA polymerase II subunit that determines amanitin sensitivity, was inserted into a bacterial plasmid cloning vehicle useful for over-production of hybrid proteins. Two plasmid const ... Link to item Cite

agr-Amanitin Tolerance in Mycophagous Drosophila.

Journal Article Science · July 8, 1983 Six species of Drosophila were tested for tolerance to the mushroom toxin alpha-amanitin, a potent inhibitor of RNA polymerase II. Three nonmycophagous species-D. melanogaster, D. immigrans, and D. pseudoobscura-showed very low survival and long developmen ... Full text Link to item Cite

Identification of a structural gene for a RNA polymerase II polypeptide in Drosophila melanogaster and mammalian species.

Journal Article Proc Natl Acad Sci U S A · June 1983 Using subclones representing 14 kilobase pairs (kb) of DNA from the Drosophila melanogaster RNA polymerase II (EC 2.7.7.6) X-linked genetic locus, RpII, we have identified four poly(A)+ RNA transcripts in adult flies. The DNA encoding only one of these, a ... Full text Link to item Cite

AMANITIN-RESISTANT RNA POLYMERASE-II MUTATIONS ARE IN THE ENZYMES LARGEST SUBUNIT

Journal Article JOURNAL OF BIOLOGICAL CHEMISTRY · January 1, 1983 Link to item Cite

A STRUCTURAL GENE FOR AN RNA POLYMERASE-II SUBUNIT

Journal Article FEDERATION PROCEEDINGS · January 1, 1983 Link to item Cite

Molecular cloning of sequences from a Drosophila RNA polymerase II locus by P element transposon tagging.

Journal Article Cell · December 1982 We have identified a lethal mutation in the D. melanogaster RNA polymerase II locus, RpIIC4, caused by insertion of a transposable element associated with the phenomenon of hybrid dysgenesis (P element). Using previously cloned P element sequences as a hyb ... Full text Link to item Cite

Immunological studies of RNA polymerase II using antibodies to subunits of Drosophila and wheat germ enzyme.

Journal Article J Biol Chem · May 25, 1982 We induced goat antibodies to Drosophila RNA polymerase II and rabbit antibodies to the isolated 215,000-dalton and 140,000-dalton polymerase II subunits (P215 and P140, respectively). Similarly, we induced rabbit antibodies to wheat germ RNA polymerase II ... Link to item Cite

Properties of mutationally altered RNA polymerases II of Drosophila.

Journal Article J Biol Chem · February 25, 1982 We tested and compared several in vitro properties of wild type and mutant RNA polymerases II from Drosophila melanogaster, using several different mutants of a single X-linked genetic locus, RpIIC4 (Greenleaf, A. L., Weeks, J. R., Voelker, R. A., Ohnishi, ... Link to item Cite

Genetic and biochemical characterization of mutants at an RNA polymerase II locus in D. melanogaster.

Journal Article Cell · October 1980 We previously described an alpha-amanitin-resistant mutant of D. melanogaster (AmaC4 or simply C4) with an altered, amanitin-resistant RNA polymerase II. We have now more fully characterized this mutant genetically and biochemically. We genetically mapped ... Full text Link to item Cite

RNA POLYMERASE-II MUTANTS OF DROSOPHILA

Journal Article FEDERATION PROCEEDINGS · January 1, 1980 Link to item Cite

Alpha-amanitin-resistant D. melanogaster with an altered RNA polymerase II.

Journal Article Cell · November 1979 Following EMS mutagenesis we recovered a mutant of D. melanogaster that grows at concentrations of alpha-amanitin lethal to wild-type. To our knowledge this mutant represents the first example of an amanitin-resistant eucaryotic organism. The amanitin resi ... Full text Link to item Cite

RNA polymerase B (or II) in heat induced puffs of Drosophila polytene chromosomes.

Journal Article Chromosoma · January 16, 1978 Using indirect immunofluorescence visualization techniques we investigated the distribution of RNA polymerase B (or II) and histone H1 at heat shock puff loci in Drosophila melanogaster polytene chromosomes at different times during and after heat shock. A ... Full text Link to item Cite

Functional organization of polytene chromosomes.

Journal Article Cold Spring Harb Symp Quant Biol · 1978 Full text Link to item Cite

Localization of RNA polymerase in polytene chromosomes of Drosophila melanogaster.

Journal Article Proc Natl Acad Sci U S A · May 1977 RNA polymerase (RNA nucleotidyltransferase) B (or II) and histone H1 of Drosophila melanogster were localized on salivary gland polytene chromosomes using the indirect immunofluorescence technique. RNA polymerase B is present almost exclusively in puffs an ... Full text Link to item Cite

Distribution of RNA polymerase on Drosophila polytene chromosomes as studied by indirect immunofluorescence.

Journal Article Chromosoma · December 16, 1976 Using indirect immunofluorescence visualization techniques we investigated the in situ distribution of RNA polymerase B on Drosophila melanogaster polytene chromosomes. The enzyme was found at many sites distributed throughout the genome in a pattern clear ... Full text Link to item Cite

RNA polymerase from sporulating Bacillus subtilis. Purification and properties of a modified form of the enzyme containing two sporulation polypeptides.

Journal Article J Biol Chem · December 25, 1975 A new form of DNA-dependent RNA polymerase termed enzyme III has been purified from sporulating cells of Bacillus subtilis. In addition to the subunits of core RNA polymerase (beta', beta, alpha, and omega), enzyme III contains sporulation-specific polypep ... Link to item Cite

RNA polymerase B from Drosophila melanogaster larvae. Purification and partial characterization.

Journal Article Eur J Biochem · December 1, 1975 A purification procedure is described by which we obtained DNA-dependent RNA polymerase B (or II) from third-instar larvae of Drosophila melanogaster in essentially pure form. The enzyme is similar to the analogous polymerases from other eukaryotes in its ... Full text Link to item Cite

Appearance of a ribonucleic acid polymerase-binding protein in asporogenous mutants of Bacillus subtilis.

Journal Article J Bacteriol · October 1973 A 70,000-dalton protein that is found in sporulating Bacillus subtilis and that binds to ribonucleic acid polymerase is present in asporogenous mutants that proceed to or beyond stage II of sporulation, but is absent from mutants blocked at stage zero. ... Full text Link to item Cite

Loss of the sigma activity of RNA polymerase of Bacillus subtilis during sporulation.

Journal Article Proc Natl Acad Sci U S A · June 1973 The activity of the sigma subunit of the RNA polymerase of Bacillus subtilis decreases markedly during the first 2 hr of sporulation. Moreover, sigma activity remains deficient throughout the sporulation process and in dormant spores. The time course of ch ... Full text Link to item Cite

Isolation of a new RNA polymerase-binding protein from sporulating Bacillus subtilis.

Journal Article Proc Natl Acad Sci U S A · February 1973 RNA polymerase was precipitated from extracts of radioactively labeled vegetative and sporulating Bacillus subtilis with antiserum prepared against vegetative core polymerase. The precipitates were solubilized and analyzed by sodium dodecyl sulfate-polyacr ... Full text Link to item Cite