![]()
|
|
||||||||
J. Biol. Chem., Vol. 276, Issue 21, 17968-17975, May 25, 2001
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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.
ATPase Activity and Multimer Formation of PilQ Protein Are
Required for Thin Pilus Biogenesis in Plasmid R64*
*
This work was supported in part by a grant from the Ministry
of Education, Science, Sports and Culture of Japan.
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.
This article has been cited by other articles:
![]() |
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] |
||||
![]() |
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] |
||||
![]() |
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] |
||||
![]() |
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] |
||||
![]() |
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] |
||||
![]() |
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] |
||||
![]() |
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] |
||||
![]() |
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 |