Biogenesis of a putative channel protein, ComEC, required for DNA uptake: membrane topology, oligomerization and formation of disulphide bonds

被引:118
作者
Draskovic, I [1 ]
Dubnau, D [1 ]
机构
[1] Int Ctr Publ Hlth, Publ Hlth Res Inst, Newark, NJ 07103 USA
关键词
D O I
10.1111/j.1365-2958.2004.04430.x
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
ComEC is a putative channel protein for DNA uptake in Bacillus subtilis and other genetically transformable bacteria. Membrane topology studies suggest a model of ComEC as a multispanning membrane protein with seven transmembrane segments (TMSs), and possibly with one laterally inserted amphipathic helix. We show that ComEC contains an intramolecular disulphide bond in its N-terminal extracellular loop (between the residues C131 and C172), which is required for the stability of the protein, and is probably introduced by BdbDC, a pair of competence-induced oxidoreductase proteins. By in vitro cross-linking using native cysteine residues we show that ComEC forms an oligomer. The oligomerization surface includes a transmembrane segment, TMS-G, near the cytoplasmic C-terminus of ComEC.
引用
收藏
页码:881 / 896
页数:16
相关论文
共 43 条
[11]   Prediction of transmembrane alpha-helices in prokaryotic membrane proteins: the dense alignment surface method [J].
Cserzo, M ;
Wallin, E ;
Simon, I ;
vonHeijne, G ;
Elofsson, A .
PROTEIN ENGINEERING, 1997, 10 (06) :673-676
[12]   EFFECT OF STAGE-0 SPORULATION MUTATIONS ON SUBTILISIN EXPRESSION [J].
FERRARI, E ;
HOWARD, SMH ;
HOCH, JA .
JOURNAL OF BACTERIOLOGY, 1986, 166 (01) :173-179
[13]   TOPOLOGICAL FRUSTRATION IN MULTISPANNING ESCHERICHIA-COLI INNER MEMBRANE-PROTEINS [J].
GAFVELIN, G ;
VONHEIJNE, G .
CELL, 1994, 77 (03) :401-412
[14]   ISOLATION AND CHARACTERIZATION OF TN917LAC-GENERATED COMPETENCE MUTANTS OF BACILLUS-SUBTILIS [J].
HAHN, J ;
ALBANO, M ;
DUBNAU, D .
JOURNAL OF BACTERIOLOGY, 1987, 169 (07) :3104-3109
[15]   CHARACTERIZATION OF COME, A LATE COMPETENCE OPERON OF BACILLUS-SUBTILIS REQUIRED FOR THE BINDING AND UPTAKE OF TRANSFORMING DNA [J].
HAHN, J ;
INAMINE, G ;
KOZLOV, Y ;
DUBNAU, D .
MOLECULAR MICROBIOLOGY, 1993, 10 (01) :99-111
[16]   Improving the predictive value of the competence transcription factor (ComK) binding site in Bacillus subtilis using a genomic approach [J].
Hamoen, LW ;
Smits, WK ;
de Jong, A ;
Holsappel, S ;
Kuipers, OP .
NUCLEIC ACIDS RESEARCH, 2002, 30 (24) :5517-5528
[17]  
Hughson AG, 1997, PROTEIN SCI, V6, P315
[18]   COMEA, A BACILLUS-SUBTILIS INTEGRAL MEMBRANE-PROTEIN REQUIRED FOR GENETIC-TRANSFORMATION, IS NEEDED FOR BOTH DNA-BINDING AND TRANSPORT [J].
INAMINE, GS ;
DUBNAU, D .
JOURNAL OF BACTERIOLOGY, 1995, 177 (11) :3045-3051
[19]   A MODEL RECOGNITION APPROACH TO THE PREDICTION OF ALL-HELICAL MEMBRANE-PROTEIN STRUCTURE AND TOPOLOGY [J].
JONES, DT ;
TAYLOR, WR ;
THORTON, JM .
BIOCHEMISTRY, 1994, 33 (10) :3038-3049
[20]   Basic charge clusters and predictions of membrane protein topology [J].
Juretic, D ;
Zoranic, L ;
Zucic, D .
JOURNAL OF CHEMICAL INFORMATION AND COMPUTER SCIENCES, 2002, 42 (03) :620-632