|
|
|
|||||||
|
|||||||||
J. Biol. Chem., Vol. 277, Issue 29, 25920-25928, July 19, 2002
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
From the Departments of The Ty3 retrovirus-like element inserts
preferentially at the transcription initiation sites of genes
transcribed by RNA polymerase III. The requirements for
transcription factor (TF) IIIC and TFIIIB in Ty3 integration
into the two initiation sites of the U6 gene carried on pU6LboxB were
previously examined. Ty3 integrates at low but detectable frequencies
in the presence of TFIIIB subunits Brf1 and TATA-binding protein.
Integration increases in the presence of the third subunit, Bdp1.
TFIIIC is not essential, but the presence of TFIIIC specifies an
orientation of TFIIIB for transcriptional initiation and directs
integration to the U6 gene-proximal initiation site. In the current
study, recombinant wild type TATA-binding protein, wild type and mutant
Brf1, and Bdp1 proteins and highly purified TFIIIC were used to
investigate the roles of specific protein domains in Ty3 integration.
The amino-terminal half of Brf1, which contains a TFIIB-like repeat,
contributed more strongly than the carboxyl-terminal half of Brf1 to
Ty3 targeting. Each half of Bdp1 split at amino acid 352 enhanced
integration. In the presence of TFIIIB and TFIIIC, the pattern of
integration extended downstream by several base pairs compared with the
pattern observed in vitro in the absence of TFIIIC and
in vivo, suggesting that TFIIIC may not be present on genes
targeted by Ty3 in vivo. Mutations in Bdp1 that affect its
interaction with TFIIIC resulted in TFIIIC-independent patterns of Ty3
integration. Brf1 zinc ribbon and Bdp1 internal deletion mutants that
are competent for polymerase III recruitment but defective in promoter
opening were competent for Ty3 integration irrespective of the state of
DNA supercoiling. These results extend the similarities between the
TFIIIB domains required for transcription and Ty3 integration and also
reveal requirements that are specific to transcription.
Mutational Analysis of the Transcription Factor IIIB-DNA Target
of Ty3 Retroelement Integration*
§¶,
, and**
Microbiology and Molecular
Genetics and ** Biological Chemistry, University of
California, Irvine, California 92697-1700 and the Division of
Biology and Center for Molecular Genetics, University of California,
San Diego, La Jolla, California 92093
*
This work was supported by United States Public Health
Service Grants GM33281 (at University of California, Irvine) and
GM18386 (at University of California, San Diego) from the National
Institutes of Health.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.
E-mail: sbsandme@uci.edu.
Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.
CiteULike 
Complore 
Connotea 
Del.icio.us 
Digg 
Reddit 
Technorati What's this?
This article has been cited by other articles:
|
|
 |
|
  X. Gao, Y. Hou, H. Ebina, H. L. Levin, and D. F. Voytas Chromodomains direct integration of retrotransposons to heterochromatin Genome Res., March 1, 2008; 18(3): 359 - 369. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 T. Chung, O. Siol, T. Dingermann, and T. Winckler Protein Interactions Involved in tRNA Gene-Specific Integration of Dictyostelium discoideum Non-Long Terminal Repeat Retrotransposon TRE5-A Mol. Cell. Biol., December 15, 2007; 27(24): 8492 - 8501. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. N. Dubey and M. R. Gartenberg A tDNA establishes cohesion of a neighboring silent chromatin domain Genes & Dev., September 1, 2007; 21(17): 2150 - 2160. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 G. A. Kassavetis, R. Driscoll, and E. P. Geiduschek Mapping the Principal Interaction Site of the Brf1 and Bdp1 Subunits of Saccharomyces cerevisiae TFIIIB J. Biol. Chem., May 19, 2006; 281(20): 14321 - 14329. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 Z. Mou, A. E. Kenny, and M. J. Curcio Hos2 and Set3 Promote Integration of Ty1 Retrotransposons at tRNA Genes in Saccharomyces cerevisiae Genetics, April 1, 2006; 172(4): 2157 - 2167. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. E. Gelbart, N. Bachman, J. Delrow, J. D. Boeke, and T. Tsukiyama Genome-wide identification of Isw2 chromatin-remodeling targets by localization of a catalytically inactive mutant Genes & Dev., April 15, 2005; 19(8): 942 - 954. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
N. Bachman, M. E. Gelbart, T. Tsukiyama, and J. D. Boeke TFIIIB subunit Bdp1p is required for periodic integration of the Ty1 retrotransposon and targeting of Isw2p to S. cerevisiae tDNAs Genes & Dev., April 15, 2005; 19(8): 955 - 964. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. Cherepanov, E. Devroe, P. A. Silver, and A. Engelman Identification of an Evolutionarily Conserved Domain in Human Lens Epithelium-derived Growth Factor/Transcriptional Co-activator p75 (LEDGF/p75) That Binds HIV-1 Integrase J. Biol. Chem., November 19, 2004; 279(47): 48883 - 48892. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
N. Bachman, Y. Eby, and J. D. Boeke Local Definition of Ty1 Target Preference by Long Terminal Repeats and Clustered tRNA Genes Genome Res., July 1, 2004; 14(7): 1232 - 1247. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 Y. Zhu, J. Dai, P. G. Fuerst, and D. F. Voytas From the Cover: Controlling integration specificity of a yeast retrotransposon PNAS, May 13, 2003; 100(10): 5891 - 5895. [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 |