JBC

HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
QUICK SEARCH:   [advanced]


     

Originally published In Press as doi:10.1074/jbc.M010652200 on March 12, 2001

J. Biol. Chem., Vol. 276, Issue 21, 17968-17975, May 25, 2001
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow All Versions of this Article:
276/21/17968    most recent
M010652200v1
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Right arrow Citation Map
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 Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Sakai, D.
Right arrow Articles by Komano, T.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Sakai, D.
Right arrow Articles by Komano, T.
Social Bookmarking
Add to CiteULike   Add to Complore   Add to Connotea   Add to Del.icio.us   Add to Digg   Add to Reddit   Add to Technorati  
What's this?

ATPase Activity and Multimer Formation of PilQ Protein Are Required for Thin Pilus Biogenesis in Plasmid R64*

Daisuke Sakai, Takayuki Horiuchi, and Teruya KomanoDagger

From the Department of Biology, Tokyo Metropolitan University, Minamiohsawa, Hachioji, Tokyo 192-0397, Japan

Plasmid R64 pilQ gene is essential for the formation of thin pilus, a type IV pilus. The pilQ product contains NTP binding motifs and belongs to the PulE-VirB11 family of NTPases. The pilQ gene was overexpressed with an N-terminal His tag, and PilQ protein was purified. Purified His tag PilQ protein displayed ATPase activity with a Vmax of 0.71 nmol/min/mg of protein and a Km of 0.26 mM at pH 6.5. By gel filtration chromatography, PilQ protein was eluted at the position corresponding to 460 kDa, suggesting that PilQ protein forms a homooctamer. To analyze the relationship between structure and function of PilQ protein, amino acid substitutions were introduced within several conserved motifs. Among 11 missense mutants, 7 mutants exhibited various levels of reduced DNA transfer frequencies in liquid matings. Four mutant genes (T234I, K238Q, D263N, and H328A) were overexpressed with a His tag. The purified mutant PilQ proteins contained various levels of reduced ATPase activity. Three mutant PilQ proteins formed stable multimers similar to wild-type PilQ, whereas the PilQ D263N multimer was unstable. PilQ D263N monomer exhibited low ATPase activity, while PilQ D263N multimer did not. These results indicate that ATPase activity of the PilQ multimer is essential for R64 thin pilus biogenesis.

* This work was supported in part by a grant from the Ministry of Education, Science, Sports and Culture of Japan.

Dagger To whom correspondence should be addressed. Tel.: 81-426-77-2568; Fax: 81-426-77-2559; E-mail: komano-teruya@c.metro-u.ac.jp.

Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.
Add to CiteULike CiteULike   Add to Complore Complore   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us   Add to Digg Digg   Add to Reddit Reddit   Add to Technorati Technorati    What's this?


This article has been cited by other articles:


Home page
 J. Bacteriol.Home page
V. Jakovljevic, S. Leonardy, M. Hoppert, and L. Sogaard-Andersen
PilB and PilT Are ATPases Acting Antagonistically in Type IV Pilus Function in Myxococcus xanthus
J. Bacteriol., April 1, 2008; 190(7): 2411 - 2421.
[Abstract] [Full Text] [PDF]

Home page
 MicrobiologyHome page
P. Chiang, L. M. Sampaleanu, M. Ayers, M. Pahuta, P. L. Howell, and L. L. Burrows
Functional role of conserved residues in the characteristic secretion NTPase motifs of the Pseudomonas aeruginosa type IV pilus motor proteins PilB, PilT and PilU
Microbiology, January 1, 2008; 154(1): 114 - 126.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
L. J. Crowther, A. Yamagata, L. Craig, J. A. Tainer, and M. S. Donnenberg
The ATPase Activity of BfpD Is Greatly Enhanced by Zinc and Allosteric Interactions with Other Bfp Proteins
J. Biol. Chem., July 1, 2005; 280(26): 24839 - 24848.
[Abstract] [Full Text] [PDF]

Home page
 J. Bacteriol.Home page
J. L. Camberg and M. Sandkvist
Molecular Analysis of the Vibrio cholerae Type II Secretion ATPase EpsE
J. Bacteriol., January 1, 2005; 187(1): 249 - 256.
[Abstract] [Full Text] [PDF]

Home page
 J. Bacteriol.Home page
J. A. Sexton, J. S. Pinkner, R. Roth, J. E. Heuser, S. J. Hultgren, and J. P. Vogel
The Legionella pneumophila PilT Homologue DotB Exhibits ATPase Activity That Is Critical for Intracellular Growth
J. Bacteriol., March 15, 2004; 186(6): 1658 - 1666.
[Abstract] [Full Text] [PDF]

Home page
 J. Bacteriol.Home page
T. J. Herdendorf, D. R. McCaslin, and K. T. Forest
Aquifex aeolicus PilT, Homologue of a Surface Motility Protein, Is a Thermostable Oligomeric NTPase
J. Bacteriol., December 1, 2002; 184(23): 6465 - 6471.
[Abstract] [Full Text] [PDF]

Home page
 J. Bacteriol.Home page
N. Bose, S. M. Payne, and R. K. Taylor
Type 4 Pilus Biogenesis and Type II-Mediated Protein Secretion by Vibrio cholerae Occur Independently of the TonB-Facilitated Proton Motive Force
J. Bacteriol., April 15, 2002; 184(8): 2305 - 2309.
[Abstract] [Full Text]

Home page
 J. Bacteriol.Home page
D. Sakai and T. Komano
Genes Required for Plasmid R64 Thin-Pilus Biogenesis: Identification and Localization of Products of the pilK, pilM, pilO, pilP, pilR, and pilT Genes
J. Bacteriol., January 15, 2002; 184(2): 444 - 451.
[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 
Copyright © 2001 by the American Society for Biochemistry and Molecular Biology.