We intend to recreate the bidirectional replication of p4 DNA in a purified, reconstituted, in vitro reaction. Satellite phage p4 DNA replicates bidirectionally, starting as a double-stranded circular molecule of 11 kb. p4 does not need the host proteins that prime the synthesis of DNA chains (dnaB, dnaG) and must synthesize its own priming apparatus. We observe p4 DNA synthesis in vitro, depending on the p4 Alpha gene product, which we have purified to near-homogeniety. The p4 Alpha protein can prime DNA synthesis on a circular, single-stranded DNA template. We shall characterize the primers made by the p4 Alpha protein, as well as any nuclease and topoisomerase activities that it may have. We shall determine the base sequence of the replication origin and show whether our in vitro replication is origin specific. Bidirectional DNA replication is a key event in the regulation of human cell growth. Understanding of the initiation of DNA replication is therefore essential for the control of normal development, wound-healing and oncogenic transformation.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Method to Extend Research in Time (MERIT) Award (R37)
Project #
5R37AI008722-22
Application #
3480476
Study Section
Microbial Physiology and Genetics Subcommittee 2 (MBC)
Project Start
1977-02-01
Project End
1992-01-31
Budget Start
1990-02-01
Budget End
1991-01-31
Support Year
22
Fiscal Year
1990
Total Cost
Indirect Cost
Name
University of California Berkeley
Department
Type
Schools of Arts and Sciences
DUNS #
094878337
City
Berkeley
State
CA
Country
United States
Zip Code
94704