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

Publication ,  Journal Article
MacKellar, AL; Greenleaf, AL
Published in: 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 bind the phosphorylated CTD and use it to localize to nascent pre-mRNA during transcription. In Saccharomyces cerevisiae, the mRNA export protein Yra1 (ALY/RNA export factor in metazoa) cotranscriptionally associates with mRNA and delivers it to the nuclear pore complex for export to the cytoplasm. Here we report that Yra1 directly binds in vitro the hyperphosphorylated form of the CTD characteristic of elongating RNA polymerase II and contains a phospho-CTD-interacting domain within amino acids 18-184, which also include an "RNA recognition motif" (RRM) (residues 77-184). Using UV cross-linking, we showed that the RRM alone binds RNA, although a larger segment extending to the C terminus (amino acids 77-226) displayed stronger RNA binding activity. Although the RRM is implicated in both RNA and CTD binding, RRM point mutations separated these two functions. Both functions are important in vivo as RNA binding-defective or CTD binding-defective versions of Yra1 engendered growth and mRNA export defects. We also report the construction and characterization of a useful new temperature-sensitive YRA1 allele (R107A/F126A). Using ChIP, we demonstrated that removing the N-terminal 76 amino acids of Yra1 (all of the phospho-CTD-interacting domain up to the RRM) results in a 10-fold decrease in Yra1 recruitment to genes during elongation. These results indicate that the phospho-CTD is likely involved directly in the cotranscriptional recruitment of Yra1.

Duke Scholars

Published In

J Biol Chem

DOI

EISSN

1083-351X

Publication Date

October 21, 2011

Volume

286

Issue

42

Start / End Page

36385 / 36395

Location

United States

Related Subject Headings

  • Transcription, Genetic
  • Saccharomyces cerevisiae Proteins
  • Saccharomyces cerevisiae
  • RNA-Binding Proteins
  • RNA, Messenger
  • RNA, Fungal
  • RNA Polymerase II
  • Protein Structure, Tertiary
  • Protein Binding
  • Phosphorylation
 

Citation

APA
Chicago
ICMJE
MLA
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MacKellar, A. L., & Greenleaf, A. L. (2011). Cotranscriptional association of mRNA export factor Yra1 with C-terminal domain of RNA polymerase II. J Biol Chem, 286(42), 36385–36395. https://doi.org/10.1074/jbc.M111.268144
MacKellar, April L., and Arno L. Greenleaf. “Cotranscriptional association of mRNA export factor Yra1 with C-terminal domain of RNA polymerase II.J Biol Chem 286, no. 42 (October 21, 2011): 36385–95. https://doi.org/10.1074/jbc.M111.268144.
MacKellar AL, Greenleaf AL. Cotranscriptional association of mRNA export factor Yra1 with C-terminal domain of RNA polymerase II. J Biol Chem. 2011 Oct 21;286(42):36385–95.
MacKellar, April L., and Arno L. Greenleaf. “Cotranscriptional association of mRNA export factor Yra1 with C-terminal domain of RNA polymerase II.J Biol Chem, vol. 286, no. 42, Oct. 2011, pp. 36385–95. Pubmed, doi:10.1074/jbc.M111.268144.
MacKellar AL, Greenleaf AL. Cotranscriptional association of mRNA export factor Yra1 with C-terminal domain of RNA polymerase II. J Biol Chem. 2011 Oct 21;286(42):36385–36395.

Published In

J Biol Chem

DOI

EISSN

1083-351X

Publication Date

October 21, 2011

Volume

286

Issue

42

Start / End Page

36385 / 36395

Location

United States

Related Subject Headings

  • Transcription, Genetic
  • Saccharomyces cerevisiae Proteins
  • Saccharomyces cerevisiae
  • RNA-Binding Proteins
  • RNA, Messenger
  • RNA, Fungal
  • RNA Polymerase II
  • Protein Structure, Tertiary
  • Protein Binding
  • Phosphorylation