REGULATION OF GENE EXPRESSION IN BACTERIOPHAGE T4

  • Mustaev, Arkady (PI)
  • Goldfarb, Alexander (PI)
  • Goldfarb, Alexander (PI)
  • Goldfarb, Alexander (PI)
  • Goldfarb, Alexander (PI)
  • MUSTAEV, ARKADY (PI)
  • MUSTAEV, ARKADY (PI)

Project Details

Description

After infection of E. coli by bacteriophage T4, the host RNA polymerase
acquires several small phage-induced polypeptides and its alfa subunits are
ADP-ribosylated. The role of these modifications in transcription control
will be studied using a combination of biochemical, genetic and
physiological approaches. We have purified four of the associated
polypeptides (15K, 19K, 25K and 29K proteins) as well as RNA polymerases
differing in the state of ADP-ribosylation and propose to study their
interactions in direct binding assays. The kinetics of in vivo formation
of these proteins and their interactions with other intracellular
components will be analysed in order to understand the coordination between
transcription and other events of T4 development such as DNA replication.
We will identify T4 genes involved in the modifications and use their
mutants to determine their role in phage development. In vitro experiments
are proposed to study the interaction of normal and modified RNA polymerase
with individual promoters with emphasis given to the change of specificity
of promoter recognition from early to late sites. We have already shown
that 25K protein induces late promoter specificity while 15K protein
decreases the utilization of early promoters. To understand the molecular
basis of the specificity change we will determine the kinetic parameters of
promoter functioning with different forms of RNA polymerase. Experiments
directed at biochemical identification of T4-induced antitermination
mechanism are also proposed. The in vitro experiments will be backed up by
the analysis of in vivo functioning of the same transcription sites using
recently sequenced tRNA gene region of T4 as the experimental system.
StatusFinished
Effective start/end date12/31/896/30/13

Funding

  • National Institute of General Medical Sciences: $351,000.00
  • National Institute of General Medical Sciences: $315,900.00
  • National Institute of General Medical Sciences: $532,510.00
  • National Institute of General Medical Sciences: $344,016.00
  • National Institute of General Medical Sciences: $347,490.00
  • National Institute of General Medical Sciences: $498,052.00
  • National Institute of General Medical Sciences: $34,796.00

ASJC

  • Medicine(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Genetics
  • Molecular Biology
  • Cell Biology

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