The overall objective of the research is to study the enzymatic initiation of DNA synthesis in mammalian cells and in particular, the mechanism of RNA priming of DNA synthesis.
The specific aims of this proposal are: 1. Examine the synthesis of RNA primers by a DNA primase that we have purified from mouse cells. 2. Examine the initiation of DNA synthesis by primase. 3. Study the mechanism of DNA initiation and elongation that may require other enzymes and/or accessory factors with well defined templates as SV40 or artificial replication forks. 4. Explore the use of anti-peptide antibody immunoaffinity methods for purification of primase and for co-purification of other proteins that may interact with primase.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM029091-06
Application #
3276584
Study Section
Molecular Biology Study Section (MBY)
Project Start
1981-06-01
Project End
1989-05-31
Budget Start
1986-06-01
Budget End
1987-05-31
Support Year
6
Fiscal Year
1986
Total Cost
Indirect Cost
Name
University of California San Diego
Department
Type
Schools of Medicine
DUNS #
077758407
City
La Jolla
State
CA
Country
United States
Zip Code
92093
Tseng, B Y; Prussak, C E (1989) Sequence and structural requirements for primase initiation in the SV40 origin of replication. Nucleic Acids Res 17:1953-63
Tseng, B Y; Prussak, C E; Almazan, M T (1989) Primase p49 mRNA expression is serum stimulated but does not vary with the cell cycle. Mol Cell Biol 9:1940-5
Prussak, C E; Tseng, B Y (1989) DNA polymerase alpha activity is not affected by protein kinases or alkaline phosphatase. Biochem Biophys Res Commun 159:1397-403
Prussak, C E; Almazan, M T; Tseng, B Y (1989) Peptide production from proteins separated by sodium dodecyl-sulfate polyacrylamide gel electrophoresis. Anal Biochem 178:233-8
Prussak, C E; Almazan, M T; Tseng, B Y (1989) Mouse primase p49 subunit molecular cloning indicates conserved and divergent regions. J Biol Chem 264:4957-63
Prussak, C E; Tseng, B Y (1987) Isolation of the DNA polymerase alpha core enzyme from mouse cells. J Biol Chem 262:6018-22