|
|
|
|||||||
|
|||||||||
J. Biol. Chem., Vol. 277, Issue 40, 37339-37348, October 4, 2002
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
From the Institut de Biologie et Chimie des Protéines, Centre
National de la Recherche Scientifique, Université de Lyon, 69367 Lyon cedex 07, France
Protein Wzc from Escherichia
coli is a member of a newly defined family of protein-tyrosine
autokinases that are essential for surface polysaccharide production in
both Gram-negative and Gram-positive bacteria. Although the catalytic
mechanism of the autophosphorylation of Wzc was recently described, the
in vivo structural organization of this protein remained
unclear. Here, we have determined the membrane topology of Wzc by
performing translational fusions of lacZ and
phoA reporter genes to the wzc gene. It has
been shown that Wzc consists of two main structural domains: an
N-terminal domain, bordered by two transmembrane helices, which is
located in the periplasm of cells, and a C-terminal domain, harboring
all phosphorylation sites of the protein, which is located in the
cytoplasm. In addition, it has been demonstrated for the first time
that Wzc can oligomerize in vivo to form essentially trimers and hexamers. Cross-linking experiments performed on strains expressing various domains of Wzc have shown that the cytoplasmic C-terminal domain is sufficient to generate oligomerization of Wzc.
Mutant proteins, modified in either the ATP-binding site or the
different phosphorylation sites, i.e. rendered unable to undergo autophosphorylation, have appeared to oligomerize into high
molecular mass species identical to those formed by the wild-type protein. It was concluded that phosphorylation of Wzc is not essential to its oligomerization. These data, connected with the
phosphorylation mechanism of Wzc, may be of biological significance in
the regulatory role played by this kinase in polysaccharide synthesis.
Structural Organization of the Protein-tyrosine Autokinase Wzc
within Escherichia coli Cells*
*
This work was supported by grants from the
Société Ezus-Lyon 1 (Contract 482.022), the Ministère
de la Recherche (Contract FNS 2000 Microbiologie), and the Institut
Universitaire de France.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: Institut de
Biologie et Chimie des Protéines, Centre National de la Recherche Scientifique, Université de Lyon, 7 Passage du Vercors, 69367 Lyon cedex 07, France. Tel.: 33-0-4-72-72-26-72; Fax:
33-0-4-72-72-26-01; E-mail: aj.cozzone@ibcp.fr.
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:
|
|
 |
|
  K. Larue, M. S. Kimber, R. Ford, and C. Whitfield Biochemical and Structural Analysis of Bacterial O-antigen Chain Length Regulator Proteins Reveals a Conserved Quaternary Structure J. Biol. Chem., March 13, 2009; 284(11): 7395 - 7403. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Marrichi, L. Camacho, D. G. Russell, and M. P. DeLisa Genetic Toggling of Alkaline Phosphatase Folding Reveals Signal Peptides for All Major Modes of Transport across the Inner Membrane of Bacteria J. Biol. Chem., December 12, 2008; 283(50): 35223 - 35235. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. F. Collins, K. Beis, C. Dong, C. H. Botting, C. McDonnell, R. C. Ford, B. R. Clarke, C. Whitfield, and J. H. Naismith The 3D structure of a periplasm-spanning platform required for assembly of group 1 capsular polysaccharides in Escherichia coli PNAS, February 13, 2007; 104(7): 2390 - 2395. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. S. Ferreira, J. H. Leitao, S. A. Sousa, A. M. Cosme, I. Sa-Correia, and L. M. Moreira Functional Analysis of Burkholderia cepacia Genes bceD and bceF, Encoding a Phosphotyrosine Phosphatase and a Tyrosine Autokinase, Respectively: Role in Exopolysaccharide Biosynthesis and Biofilm Formation Appl. Envir. Microbiol., January 1, 2007; 73(2): 524 - 534. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. F. Collins, K. Beis, B. R. Clarke, R. C. Ford, M. Hulley, J. H. Naismith, and C. Whitfield Periplasmic Protein-Protein Contacts in the Inner Membrane Protein Wzc Form a Tetrameric Complex Required for the Assembly of Escherichia coli Group 1 Capsules J. Biol. Chem., January 27, 2006; 281(4): 2144 - 2150. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. Peleg, Y. Shifrin, O. Ilan, C. Nadler-Yona, S. Nov, S. Koby, K. Baruch, S. Altuvia, M. Elgrably-Weiss, C. M. Abe, et al. Identification of an Escherichia coli Operon Required for Formation of the O-Antigen Capsule J. Bacteriol., August 1, 2005; 187(15): 5259 - 5266. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. N. Reid and C. Whitfield Functional Analysis of Conserved Gene Products Involved in Assembly of Escherichia coli Capsules and Exopolysaccharides: Evidence for Molecular Recognition between Wza and Wzc for Colanic Acid Biosynthesis J. Bacteriol., August 1, 2005; 187(15): 5470 - 5481. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Nesper, C. M. D. Hill, A. Paiment, G. Harauz, K. Beis, J. H. Naismith, and C. Whitfield Translocation of Group 1 Capsular Polysaccharide in Escherichia coli Serotype K30: STRUCTURAL AND FUNCTIONAL ANALYSIS OF THE OUTER MEMBRANE LIPOPROTEIN Wza J. Biol. Chem., December 12, 2003; 278(50): 49763 - 49772. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. Grangeasse, B. Obadia, I. Mijakovic, J. Deutscher, A. J. Cozzone, and P. Doublet Autophosphorylation of the Escherichia coli Protein Kinase Wzc Regulates Tyrosine Phosphorylation of Ugd, a UDP-glucose Dehydrogenase J. Biol. Chem., October 10, 2003; 278(41): 39323 - 39329. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. K. Morona, R. Morona, D. C. Miller, and J. C. Paton Mutational Analysis of the Carboxy-Terminal (YGX)4 Repeat Domain of CpsD, an Autophosphorylating Tyrosine Kinase Required for Capsule Biosynthesis in Streptococcus pneumoniae J. Bacteriol., May 15, 2003; 185(10): 3009 - 3019. [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 |