Abstract

Through the use of natural template probes many components of the E. coli replicative machinery have been isolated. These components include a priming enzyme, a single-stranded DNA binding protein and the complex DNA polymerase III holoenzyme that consists of at least seven different subunits. The functional role of many replication proteins is becoming understood; however, little is known of the regulation and coordination of their synthesis. This project proposes to investigate this regulatory problem by using recombinant plasmids (i) to increase in vivo the concentration of a specific replication gene and (ii) to direct an in vitro protein synthesizing system. The effect of gene dosage upon other replication gene products will be examined by functional and immunological assays. This may detect coordinate relationships between replication protein levels. An in vitro protein synthesizing system may (i) permit identification of the protein product of a given dna gene, (ii) allow detection of precursors to active replication proteins, and (iii) provide an assay for specific effectors of replication protein transcription or translation.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM036255-02
Application #
3289823
Study Section
Biochemistry Study Section (BIO)
Project Start
1985-09-01
Project End
1987-08-31
Budget Start
1986-09-01
Budget End
1987-08-31
Support Year
2
Fiscal Year
1986
Total Cost
Indirect Cost

  Institution

Name
University of Colorado Denver
Department
Type
Schools of Medicine
DUNS #
065391526
City
Aurora
State
CO
Country
United States
Zip Code
80045

  Publications

Muthuswami, Rohini; Chen, Joe; Burnett, Bruce P et al. (2002) The HIV plus-strand transfer reaction: determination of replication-competent intermediates and identification of a novel lentiviral element, the primer over-extension sequence. J Mol Biol 315:311-23
Glover, B P; McHenry, C S (2001) The DNA polymerase III holoenzyme: an asymmetric dimeric replicative complex with leading and lagging strand polymerases. Cell 105:925-34
Gao, D; McHenry, C S (2001) tau binds and organizes Escherichia coli replication proteins through distinct domains. Domain IV, located within the unique C terminus of tau, binds the replication fork, helicase, DnaB. J Biol Chem 276:4441-6
Dallmann, H G; Kim, S; Pritchard, A E et al. (2000) Characterization of the unique C terminus of the Escherichia coli tau DnaX protein. Monomeric C-tau binds alpha AND DnaB and can partially replace tau in reconstituted replication forks. J Biol Chem 275:15512-9
Marians, K J; Hiasa, H; Kim, D R et al. (1998) Role of the core DNA polymerase III subunits at the replication fork. Alpha is the only subunit required for processive replication. J Biol Chem 273:2452-7
Kim, S; Dallmann, H G; McHenry, C S et al. (1996) Tau protects beta in the leading-strand polymerase complex at the replication fork. J Biol Chem 271:4315-8
Kim, S; Dallmann, H G; McHenry, C S et al. (1996) Coupling of a replicative polymerase and helicase: a tau-DnaB interaction mediates rapid replication fork movement. Cell 84:643-50
Kim, S; Dallmann, H G; McHenry, C S et al. (1996) tau couples the leading- and lagging-strand polymerases at the Escherichia coli DNA replication fork. J Biol Chem 271:21406-12
Carter, J R; Franden, M A; Aebersold, R et al. (1993) Identification, isolation, and characterization of the structural gene encoding the delta' subunit of Escherichia coli DNA polymerase III holoenzyme. J Bacteriol 175:3812-22
Carter, J R; Franden, M A; Aebersold, R et al. (1993) Identification, isolation, and overexpression of the gene encoding the psi subunit of DNA polymerase III holoenzyme. J Bacteriol 175:5604-10

Showing the most recent 10 out of 19 publications