A two-hybrid system based on chimeric operator recognition for studying protein homo/heterodimerization in Escherichia coli

被引:32
作者
Di Lallo, G
Castagnoli, L
Ghelardini, P
Paolozzi, L
机构
[1] Univ Roma Tor Vergata, Dipartimento Biol, I-00133 Rome, Italy
[2] CNR, Ctr Acidi Nucl, Rome, Italy
来源
MICROBIOLOGY-SGM | 2001年 / 147卷
关键词
protein-protein interaction; cell division proteins; phage repressor; beta-galactosidase assay;
D O I
10.1099/00221287-147-6-1651
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
The development of a convenient and promising alternative to the various two-hybrid methods that are used to study protein-protein interactions is described. In this system, a lambdoid chimeric operator is recognized by a hybrid repressor formed by two chimeric monomers whose C-terminal domains are composed of heterologous proteins (or protein domains). Only if these proteins efficiently dimerize in vivo is a functional repressor formed able to bind the chimeric operator and shut off the synthesis of a downstream reporter gene, This new approach was tested with several interacting proteins ranging in size from less than 100 to more than 800 amino acids and, to date, no size or topology limit has been detected.
引用
收藏
页码:1651 / 1656
页数:6
相关论文
共 19 条
[1]   PEMBL - A NEW FAMILY OF SINGLE STRANDED PLASMIDS [J].
DENTE, L ;
CESARENI, G ;
CORTESE, R .
NUCLEIC ACIDS RESEARCH, 1983, 11 (06) :1645-1655
[2]   Two-hybrid assay:: construction of an Escherichia coli system to quantify homodimerization ability in vivo [J].
Di Lallo, G ;
Ghelardini, P ;
Paolozzi, L .
MICROBIOLOGY-UK, 1999, 145 :1485-1490
[3]   FtsZ dimerization in vivo [J].
Di Lallo, G ;
Anderluzzi, D ;
Ghelardini, P ;
Paolozzi, L .
MOLECULAR MICROBIOLOGY, 1999, 32 (02) :265-274
[4]   Activation of prokaryotic transcription through arbitrary protein-protein contacts [J].
Dove, SL ;
Joung, JK ;
Hochschild, A .
NATURE, 1997, 386 (6625) :627-630
[5]   A NOVEL GENETIC SYSTEM TO DETECT PROTEIN PROTEIN INTERACTIONS [J].
FIELDS, S ;
SONG, OK .
NATURE, 1989, 340 (6230) :245-246
[6]  
Hanes J, 1999, CURR TOP MICROBIOL, V243, P107
[7]   SEQUENCE REQUIREMENTS FOR COILED-COILS - ANALYSIS WITH LAMBDA-REPRESSOR-GCN4 LEUCINE ZIPPER FUSIONS [J].
HU, JC ;
OSHEA, EK ;
KIM, PS ;
SAUER, RT .
SCIENCE, 1990, 250 (4986) :1400-1403
[8]   Analysis of MinC reveals two independent domains involved in interaction with MinD and FtsZ [J].
Hu, ZL ;
Lutkenhaus, J .
JOURNAL OF BACTERIOLOGY, 2000, 182 (14) :3965-3971
[9]   Bacterial two-hybrid system that exploits a cAMP signaling cascade in Escherichia coli [J].
Karimova, G ;
Ullmann, A ;
Ladant, D .
APPLICATIONS OF CHIMERIC GENES AND HYBRID PROTEINS, PT C, 2000, 328 :59-73
[10]   A bacterial two-hybrid system based on a reconstituted signal transduction pathway [J].
Karimova, G ;
Pidoux, J ;
Ullmann, A ;
Ladant, D .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1998, 95 (10) :5752-5756