Affiliation:
1. Department of Biochemistry and Molecular Biology and Center for Microbial Structural Biology, 205 South Frear Laboratory, The Pennsylvania State University, University Park, Pennsylvania 16802
Abstract
ABSTRACT
The roles of three TATA binding protein (TBP) homologs (TBP1, TBP2, and TBP3) in the archaeon
Methanosarcina acetivorans
were investigated by using genetic and molecular approaches. Although
tbp2
and
tbp3
deletion mutants were readily obtained, a
tbp1
mutant was not obtained, and the growth of a conditional
tbp1
expression strain was tetracycline dependent, indicating that TBP1 is essential. Transcripts of
tbp1
were 20-fold more abundant than transcripts of
tbp2
and 100- to 200-fold more abundant than transcripts of
tbp3
, suggesting that TBP1 is the primary TBP utilized during growth. Accordingly,
tbp1
is strictly conserved in the genomes of
Methanosarcina
species. Δ
tbp3
and Δ
tbp2
strains exhibited an extended lag phase compared with the wild type, although the lag phase for the Δ
tbp2
strain was less pronounced when this strain was transitioning from growth on methylotrophic substrates to growth on acetate. Acetate-adapted Δ
tbp3
cells exhibited growth rates, final growth yields, and lag times that were significantly reduced compared with those of the wild type when the organisms were cultured with growth-limiting concentrations of acetate, and the acetate-adapted Δ
tbp2
strain exhibited a final growth yield that was reduced compared with that of the wild type when the organisms were cultured with growth-limiting acetate concentrations. DNA microarray analyses identified 92 and 77 genes with altered transcription in the Δ
tbp2
and Δ
tbp3
strains, respectively, which is consistent with a role for TBP2 and TBP3 in optimizing gene expression. Together, the results suggest that TBP2 and TBP3 are required for efficient growth under conditions similar to the conditions in the native environment of
M. acetivorans
.
Publisher
American Society for Microbiology
Subject
Molecular Biology,Microbiology
Cited by
16 articles.
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