| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
|
|
|
|||||||
|
|||||||||
J. Biol. Chem., Vol. 277, Issue 48, 46433-46441, November 29, 2002
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
From the McArdle Laboratory for Cancer Research, University of
Wisconsin, Madison, Wisconsin 53706
Prokaryotic RNA polymerase holoenzyme is composed
of core subunits (
Conformational Flexibility in
70 Region
2 during Transcription Initiation*
2
'
) plus a 
factor
that confers promoter specificity allowing for regulation of gene
expression. Holoenzyme is known to undergo several conformational
changes during the multiple steps of transcription initiation. However,
the effects of these changes on the functions of specific regions have
not been well characterized. In this work, we addressed the role of
possible conformational change in region 2 of Escherichia
coli 70 by engineering disulfide bonds that
"lock" region 2.1 with region 2.2 and region 2.2 with region 2.3. When these mutant holoenzymes were characterized for gross defects in
multiple-round transcription, we found that insertion of either
disulfide bond did not result in a fundamental block, indicating that
the disulfide-containing holoenzymes are active. However, both
disulfide-containing holoenzymes exhibited defects in formation and
stability of the open complex. Our results suggest that conformational
flexibility within 70 region 2 facilitates open complex
formation and transcription initiation.
*
This work was supported by Grant GM28575 from the National
Institutes of Health (to R. R. B.).The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
To whom correspondence should be addressed: University of
Wisconsin-Madison, McArdle Laboratory for Cancer Research,
1400 University Ave., Madison, WI 53706-1599. Tel.:
608-263-2635; Fax: 608-262-2824; E-mail:
burgess@oncology.wisc.edu.
CiteULike 
Complore 
Connotea 
Del.icio.us 
Digg 
Reddit 
Technorati What's this?
This article has been cited by other articles:
|
|
 |
|
  N. Barinova, E. Zhilina, I. Bass, V. Nikiforov, and A. Kulbachinskiy Lineage-Specific Amino Acid Substitutions in Region 2 of the RNA Polymerase {sigma} Subunit Affect the Temperature of Promoter Opening J. Bacteriol., April 15, 2008; 190(8): 3088 - 3092. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. K. Sorenson and S. A. Darst Disulfide cross-linking indicates that FlgM-bound and free {sigma}28 adopt similar conformations PNAS, November 7, 2006; 103(45): 16722 - 16727. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. Chakraborty and V. Nagaraja Dual Role for Transactivator Protein C in Activation of mom Promoter of Bacteriophage Mu J. Biol. Chem., March 31, 2006; 281(13): 8511 - 8517. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. Wong, G. A. Kassavetis, J.-P. Leonetti, and E. P. Geiduschek Mutational and Functional Analysis of a Segment of the Sigma Family Bacteriophage T4 Late Promoter Recognition Protein gp55 J. Biol. Chem., February 21, 2003; 278(9): 7073 - 7080. [Abstract] [Full Text] [PDF]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
| All ASBMB Journals | Molecular and Cellular Proteomics |
| Journal of Lipid Research | ASBMB Today |
