Gene Expression in Heterospecific Environments

Cohen, Stanley N.

Abstract

The proposed work is a multi-faceted project aimed at elucidating the factors that affect gene expression in heterospecific environments. Its specific objectives are: 1) isolation and characterization of transcriptional start signals from selected prokaryotic and eukaryotic sources, 2) investigation of the relationship between the strength of the transcriptional start and the level of expression of structural genes distal to the promoter, 3) use of clones and characterized promoters to express genes in heterospecific environments, 4) investigation of the structural and positional features of ribosomal binding sequences on heterospecific gene expression, 5) study of specific alterations in translational control sites on gene expression, 6) study of the role of intervening sequences of eukaryotic genomic DNA in the production of translational products with this DNA versus corresponding cDNA segments, 7) investigation of the relationship between the location of intervening sequences and the structure of multi-component peptides that are expressed as a single precursor molecule. The proposed work is made possible by recently developed methods for the manipulation of cloning DNA segments and for the detailed analysis of DNA sequence at the nucleotide level.

Funding Agency

• Agency: National Institute of Health (NIH)
• Institute: National Institute of General Medical Sciences (NIGMS)
• Type: Research Project (R01)
• Project #: 3R01GM027241-05S1
• Application #: 3274632
• Study Section: (SSS)
• Project Start: 1979-12-01
• Project End: 1986-11-30
• Budget Start: 1985-09-26
• Budget End: 1986-11-30
• Support Year: 5
• Fiscal Year: 1985

Institution

• Name: Stanford University
• Type: Schools of Medicine
• DUNS #: 800771545
• City: Stanford
• State: CA
• Country: United States
• Zip Code: 94305

Publications

Claverie-Martin, F; Wang, M; Cohen, S N (1997) ARD-1 cDNA from human cells encodes a site-specific single-strand endoribonuclease that functionally resembles Escherichia coli RNase E. J Biol Chem 272:13823-8

McDowall, K J; Cohen, S N (1996) The N-terminal domain of the rne gene product has RNase E activity and is non-overlapping with the arginine-rich RNA-binding site. J Mol Biol 255:349-55

McDowall, K J; Lin-Chao, S; Cohen, S N (1994) A+U content rather than a particular nucleotide order determines the specificity of RNase E cleavage. J Biol Chem 269:10790-6

Lin-Chao, S; Wong, T T; McDowall, K J et al. (1994) Effects of nucleotide sequence on the specificity of rne-dependent and RNase E-mediated cleavages of RNA I encoded by the pBR322 plasmid. J Biol Chem 269:10797-803

Wang, M; Cohen, S N (1994) ard-1: a human gene that reverses the effects of temperature-sensitive and deletion mutations in the Escherichia coli rne gene and encodes an activity producing RNase E-like cleavages. Proc Natl Acad Sci U S A 91:10591-5

McDowall, K J; Hernandez, R G; Lin-Chao, S et al. (1993) The ams-1 and rne-3071 temperature-sensitive mutations in the ams gene are in close proximity to each other and cause substitutions within a domain that resembles a product of the Escherichia coli mre locus. J Bacteriol 175:4245-9

Xu, F; Lin-Chao, S; Cohen, S N (1993) The Escherichia coli pcnB gene promotes adenylylation of antisense RNAI of ColE1-type plasmids in vivo and degradation of RNAI decay intermediates. Proc Natl Acad Sci U S A 90:6756-60

Klug, G; Cohen, S N (1991) Effects of translation on degradation of mRNA segments transcribed from the polycistronic puf operon of Rhodobacter capsulatus. J Bacteriol 173:1478-84

Lin-Chao, S; Cohen, S N (1991) The rate of processing and degradation of antisense RNAI regulates the replication of ColE1-type plasmids in vivo. Cell 65:1233-42

Klug, G; Cohen, S N (1990) Combined actions of multiple hairpin loop structures and sites of rate-limiting endonucleolytic cleavage determine differential degradation rates of individual segments within polycistronic puf operon mRNA. J Bacteriol 172:5140-6