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Identification of a negative regulatory element involved in tissue-specific expression of mouse renin genes.

Publication ,  Journal Article
Barrett, G; Horiuchi, M; Paul, M; Pratt, RE; Nakamura, N; Dzau, VJ
Published in: Proc Natl Acad Sci U S A
February 1, 1992

The 5' flanking region of the mouse renin genes (Ren-1d and Ren-2d) contains two motifs that are homologous to known negative regulatory elements (NREs). Ren-2d has a 150-base-pair (bp) insertion 5' to the upstream putative NRE (NRE-1), which is lacking in Ren-1d. We tested the functionality of these sequences by using site-directed mutagenesis to delete individually each putative NRE from Ren-1d and to delete the 150-bp insertion from Ren-2d. We examined the effect of these mutations on the expression of the reporter gene chloramphenicol acetyltransferase, which was expressed from a truncated thymidine kinase promoter fused to the renin regulatory region. This plasmid was transfected into human choriocarcinoma JEG-3 cells. Only the upstream NRE (positions -619 to -597) was found to be functional in Ren-1d. The deletion of a 150-bp insertion from Ren-2d resulted in the suppression of chloramphenicol acetyltransferase activity to the level of Ren-1d expression. These data suggest that the upstream NRE that is functional in Ren-1d, but not in Ren-2d, may be partly responsible for differential expression of the renin genes in various tissues. The molecular mechanism of the NRE was examined by studying its interaction with nuclear proteins in submandibular gland and JEG-3 cells by gel-mobility-shift assays. Specific nuclear protein binding was observed only to the upstream NRE and the molecular mass of this protein was approximately 72 kDa as determined by Southwestern blot analysis. Thus our results suggest that both Ren-1d and Ren-2d conserve a cis-acting NRE in the 5' flanking region. In Ren-1d, this NRE could bind a specific nuclear protein resulting in the inhibition of Ren-1d expression in these tissues. On the other hand, the NRE in Ren-2d is nonfunctional due to interference by an adjacent 150-bp insertion.

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Published In

Proc Natl Acad Sci U S A

DOI

ISSN

0027-8424

Publication Date

February 1, 1992

Volume

89

Issue

3

Start / End Page

885 / 889

Location

United States

Related Subject Headings

  • Transcription, Genetic
  • Sequence Alignment
  • Renin
  • Regulatory Sequences, Nucleic Acid
  • Oligodeoxyribonucleotides
  • Nuclear Proteins
  • Molecular Sequence Data
  • Mice
  • Gene Expression Regulation
  • DNA-Binding Proteins
 

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Barrett, G., Horiuchi, M., Paul, M., Pratt, R. E., Nakamura, N., & Dzau, V. J. (1992). Identification of a negative regulatory element involved in tissue-specific expression of mouse renin genes. Proc Natl Acad Sci U S A, 89(3), 885–889. https://doi.org/10.1073/pnas.89.3.885
Barrett, G., M. Horiuchi, M. Paul, R. E. Pratt, N. Nakamura, and V. J. Dzau. “Identification of a negative regulatory element involved in tissue-specific expression of mouse renin genes.Proc Natl Acad Sci U S A 89, no. 3 (February 1, 1992): 885–89. https://doi.org/10.1073/pnas.89.3.885.
Barrett G, Horiuchi M, Paul M, Pratt RE, Nakamura N, Dzau VJ. Identification of a negative regulatory element involved in tissue-specific expression of mouse renin genes. Proc Natl Acad Sci U S A. 1992 Feb 1;89(3):885–9.
Barrett, G., et al. “Identification of a negative regulatory element involved in tissue-specific expression of mouse renin genes.Proc Natl Acad Sci U S A, vol. 89, no. 3, Feb. 1992, pp. 885–89. Pubmed, doi:10.1073/pnas.89.3.885.
Barrett G, Horiuchi M, Paul M, Pratt RE, Nakamura N, Dzau VJ. Identification of a negative regulatory element involved in tissue-specific expression of mouse renin genes. Proc Natl Acad Sci U S A. 1992 Feb 1;89(3):885–889.
Journal cover image

Published In

Proc Natl Acad Sci U S A

DOI

ISSN

0027-8424

Publication Date

February 1, 1992

Volume

89

Issue

3

Start / End Page

885 / 889

Location

United States

Related Subject Headings

  • Transcription, Genetic
  • Sequence Alignment
  • Renin
  • Regulatory Sequences, Nucleic Acid
  • Oligodeoxyribonucleotides
  • Nuclear Proteins
  • Molecular Sequence Data
  • Mice
  • Gene Expression Regulation
  • DNA-Binding Proteins