Abstract

Dihydrofolate reductase (DHFR) gene amplification mutants will be used as model systems to identify and characterize a transcription unit whch gives rise to at least seven discrete mRNAs. A major effort will be devoted to the functional analysis of RNA polymerase II transcription termination sites. Proposed research will also identify and characterize additional trancription units or other genetic information, located in the vicinity of the DHFR gene. Such information should result in a better understanding of genome organization and the regulation of gene expression in mammalian cells. Proposed experiments are centered around the three topics listed below and will be designed to answer the specific questions listed under each topic. I. Identification and characterization of the DHFR transcription termination site. 1. Where is the DHFR transcription termination site located? 2. Does transcription termination occur at a unique site? 3. What are the DNA sequence requirements for a functional transcription termination site? II. The use of modular transcription units to study mRNA metabolism. 1. Does transcription termination play a role in gene expression? 2. What is the fate of a transcript that lacks a polyA tail? 3. What is the fate of a transcript that lacks a 5' cap structure? III. Structural and functional analysis of a transcription unit located in front of the DHFR gene. 1. What are the boundaries of this transcription unit? 2. What is the functional relationship between DHFR and this adjacent transcription unit?

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
2R01GM030204-04
Application #
3277836
Study Section
Molecular Biology Study Section (MBY)
Project Start
1982-03-01
Project End
1987-02-28
Budget Start
1985-03-01
Budget End
1986-02-28
Support Year
4
Fiscal Year
1985
Total Cost
Indirect Cost

  Institution

Name
Baylor College of Medicine
Department
Type
Schools of Medicine
DUNS #
074615394
City
Houston
State
TX
Country
United States
Zip Code
77030

  Publications

Kash, S F; Kellems, R E (1994) Control of transcription arrest in intron 1 of the murine adenosine deaminase gene. Mol Cell Biol 14:6198-207
Kulkarni, A D; Rudolph, F B; Van Buren, C T (1994) The role of dietary sources of nucleotides in immune function: a review. J Nutr 124:1442S-1446S
Winston, J H; Hong, L; Akroyd, S et al. (1994) Diverse genetic regulatory elements are required to direct the proper tissue-specific and developmental expression of the murine adenosine deaminase gene. Adv Exp Med Biol 370:579-84
Kash, S F; Innis, J W; Jackson, A U et al. (1993) Functional analysis of a stable transcription arrest site in the first intron of the murine adenosine deaminase gene. Mol Cell Biol 13:2718-29
Mohamedali, K A; Guicherit, O M; Kellems, R E et al. (1993) The highest levels of purine catabolic enzymes in mice are present in the proximal small intestine. J Biol Chem 268:23728-33
Winston, J H; Hanten, G R; Overbeek, P A et al. (1992) 5' flanking sequences of the murine adenosine deaminase gene direct expression of a reporter gene to specific prenatal and postnatal tissues in transgenic mice. J Biol Chem 267:13472-9
Innis, J W; Moore, D J; Kash, S F et al. (1991) The murine adenosine deaminase promoter requires an atypical TATA box which binds transcription factor IID and transcriptional activity is stimulated by multiple upstream Sp1 binding sites. J Biol Chem 266:21765-72
Innis, J W; Kellems, R E (1991) A heat-labile factor promotes premature 3' end formation in exon 1 of the murine adenosine deaminase gene in a cell-free transcription system. Mol Cell Biol 11:5398-409
Guicherit, O M; Rudolph, F B; Kellems, R E et al. (1991) Molecular cloning and expression of a mouse muscle cDNA encoding adenylosuccinate synthetase. J Biol Chem 266:22582-7
Mifflin, R C; Kellems, R E (1991) Coupled transcription-polyadenylation in a cell-free system. J Biol Chem 266:19593-8

Showing the most recent 10 out of 19 publications