Abstract

This project is a comprehensive study, using primarily genetic and molecular genetic methods, of the process of entry of phage T7 DNA into infected cells. The PI has developed an assay, based on the time of methylation of GATC sequences (that have been placed at specific locations on phage DNA) by dam methylase. Under normal conditions about 1 kb of phage DNA enters and is transcribed before the remainder of the DNA enters the cytoplasm. Both cis- and trans-acting phage mutants will be obtained to identify DNA sites and phage proteins that participate in this process. E. coli mutants, that absorb T7 but are not killed, will be isolated to identify potential host components of the system. The phage proteins that enter the cell with DNA will be identified and the initial locations of both the DNA and the proteins will be determined. Initial attempts will be made study the binding of phage proteins to the leading end of phage DNA. Other objectives are a preliminary analysis of the energetics of DNA translocation and characterization of the sequence on phage T5 that is responsible for arrest of translocation of T5 DNA.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM032095-16
Application #
6018575
Study Section
Microbial Physiology and Genetics Subcommittee 2 (MBC)
Project Start
1982-04-01
Project End
2000-06-30
Budget Start
1999-07-01
Budget End
2000-06-30
Support Year
16
Fiscal Year
1999
Total Cost
Indirect Cost

  Institution

Name
University of Texas Austin
Department
Biology
Type
Schools of Arts and Sciences
DUNS #
City
Austin
State
TX
Country
United States
Zip Code
78712

  Publications

Molineux, Ian J; Panja, Debabrata (2013) Popping the cork: mechanisms of phage genome ejection. Nat Rev Microbiol 11:194-204
Casjens, Sherwood R; Molineux, Ian J (2012) Short noncontractile tail machines: adsorption and DNA delivery by podoviruses. Adv Exp Med Biol 726:143-79
Nguyen, Andre H; Molineux, Ian J; Springman, Rachael et al. (2012) Multiple genetic pathways to similar fitness limits during viral adaptation to a new host. Evolution 66:363-74
Bull, J J; Vimr, E R; Molineux, I J (2010) A tale of tails: Sialidase is key to success in a model of phage therapy against K1-capsulated Escherichia coli. Virology 398:79-86
Lee, Young-Sam; Lee, Sujin; Demeler, Borries et al. (2010) Each monomer of the dimeric accessory protein for human mitochondrial DNA polymerase has a distinct role in conferring processivity. J Biol Chem 285:1490-9
Panja, Debabrata; Molineux, Ian J (2010) Dynamics of bacteriophage genome ejection in vitro and in vivo. Phys Biol 7:045006
Lee, Young-Sam; Johnson, Kenneth A; Molineux, Ian J et al. (2010) A single mutation in human mitochondrial DNA polymerase Pol gammaA affects both polymerization and proofreading activities of only the holoenzyme. J Biol Chem 285:28105-16
Savalia, Dhruti; Robins, William; Nechaev, Sergei et al. (2010) The role of the T7 Gp2 inhibitor of host RNA polymerase in phage development. J Mol Biol 402:118-26
Chang, Chung-Yu; Kemp, Priscilla; Molineux, Ian J (2010) Gp15 and gp16 cooperate in translocating bacteriophage T7 DNA into the infected cell. Virology 398:176-86
Keller, Thomas E; Molineux, Ian J; Bull, James J (2009) Viral resistance evolution fully escapes a rationally designed lethal inhibitor. Mol Biol Evol 26:2041-6

Showing the most recent 10 out of 52 publications