Skip Navigation

This Article
Right arrow Print PDF (3003K)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to My Personal Archive
Right arrow Download to citation manager
Right arrow Search for citing articles in:
ISI Web of Science (15)
Right arrow Commercial Re-use Guidelines
for Open Access NAR Content
Google Scholar
Right arrow Articles by Filutowicz, M.
Right arrow Articles by Levchenko, I.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Filutowicz, M.
Right arrow Articles by Levchenko, I.
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

Nucleic Acids Research, 1994, Vol. 22, No. 20 4211-4215
© 1994

MOLECULAR BIOLOGY

Cooperative binding of initiator protein to replication origin conferred by single amino acid substitution

Marcin Filutowicz*, Dona York and Igor Levchenko

Department of Bacteriology, University of Wisconsin Madison, Wl 53706, USA

*To whom correspondence should be addressed

Received June 17, 1994. Revised August 21, 1994. Accepted August 21, 1994.

The replication initiator protein {pi} of plasmid R6K binds seven 22 bp direct repeats (DR) in the {pi} origin. The {gamma} protein also binds to an inverted repeat (IR) in the operator of its own gene, pir, which lies outside the {gamma} origin sequences. A genetic system was devised to select for {pi} protein mutants which discriminate between IR and OR (York (et al., Gene (Amst.) 116, 7–12, 1992; York and Filutowicz, J. Biol. Chem. 268, 21854–21861, 1993). From this selection the mutant {pi}S87N protein was isolated which is deficient in repressing the pir gene's expression because it cannot bind to IR at the pir gene operator. Remarkably, we discovered that {pi}S87N binds to DR cooperatively under conditions where wt {pi} binds independently. Moreover, the {pi}S87N is more active as a replication initiator in vivo when supplied at the same level as wt {pi}. Quantitative binding assays showed that both wt {pi} and TTS87N bind a DNA fragment containing a single DR unit with a similar affinity (Kd = 0.3x10–12 M). Thus, cooperativity of {pi}TS87N is most likely achieved through altered interactions between protomers bound at adjacent DR units.


Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us    What's this?


This article has been cited by other articles:


Home page
 Proc. Natl. Acad. Sci. USAHome page
D. Lebrecht, M. Foehr, E. Smith, F. J. P. Lopes, C. E. Vanario-Alonso, J. Reinitz, D. S. Burz, and S. D. Hanes
Bicoid cooperative DNA binding is critical for embryonic patterning in Drosophila
PNAS, September 13, 2005; 102(37): 13176 - 13181.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
S. Kunnimalaiyaan, R. Kruger, W. Ross, S. A. Rakowski, and M. Filutowicz
Binding Modes of the Initiator and Inhibitor Forms of the Replication Protein {pi} to the {gamma} ori Iteron of Plasmid R6K
J. Biol. Chem., September 24, 2004; 279(39): 41058 - 41066.
[Abstract] [Full Text] [PDF]

Home page
 J. Bacteriol.Home page
R. Krüger and M. Filutowicz
Dimers of pi Protein Bind the A+T-Rich Region of the R6K gamma Origin near the Leading-Strand Synthesis Start Sites: Regulatory Implications
J. Bacteriol., May 1, 2000; 182(9): 2461 - 2467.
[Abstract] [Full Text]

Home page
 Proc. Natl. Acad. Sci. USAHome page
J. Wu, M. Sektas, D. Chen, and M. Filutowicz
Two forms of replication initiator protein: Positive and negative controls
PNAS, December 9, 1997; 94(25): 13967 - 13972.
[Abstract] [Full Text] [PDF]


Disclaimer:
Please note that abstracts for content published before 1996 were created through digital scanning and may therefore not exactly replicate the text of the original print issues. All efforts have been made to ensure accuracy, but the Publisher will not be held responsible for any remaining inaccuracies. If you require any further clarification, please contact our Customer Services Department.