Studies of the distribution of Escherichia coli cAMP-receptor protein and RNA polymerase along the E-coli chromosome

被引:232
作者
Grainger, DC [1 ]
Hurd, D
Harrison, M
Holdstock, J
Busby, SJW
机构
[1] Univ Birmingham, Sch Biosci, Birmingham B15 2TT, W Midlands, England
[2] Oxford Gene Technol, Oxford OX5 1PF, England
基金
英国惠康基金;
关键词
chromatin immunoprecipitation; genomics; stringent response; transcription;
D O I
10.1073/pnas.0506687102
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
Chromatin immunoprecipitation and high-density microarrays have been used to monitor the distribution of the global transcription regulator Escherichia coli cAMP-receptor protein (CRP) and RNA polymerase along the E. coli chromosome. Our results identify targets occupied by CRP and genes transcribed by RNA polymerase in vivo. Many of the loci of CRIP binding are at known CRIP regulated promoters. However, our results show that CRP also interacts with thousands of weaker sites across the whole chromosome and that this "background" binding can be used as a probe for organization within the E. coli folded chromosome. In rapidly growing cells, we show that the major sites of RNA polymerase binding are approximate to 90 transcription units that include genes needed for protein synthesis. Upon the addition of rifampicin, RNA polymerase is distributed among >500 functional promoters. We show that the chromatin immunoprecipitation and high-density-microarrays methodology can be used to study the redistribution of RNA polymerase induced by environmental stress, revealing previously uncharacterized aspects of RNA polymerase behavior and providing an alternative to the "transcriptomics" approach for studying global transcription patterns.
引用
收藏
页码:17693 / 17698
页数:6
相关论文
共 27 条
[1]   Evolution of transcription factors and the gene regulatory network in Escherichia coli [J].
Babu, MM ;
Teichmann, SA .
NUCLEIC ACIDS RESEARCH, 2003, 31 (04) :1234-1244
[2]   Spatial arrangement and macrodomain organization of bacterial chromosomes [J].
Boccard, F ;
Esnault, E ;
Valens, M .
MOLECULAR MICROBIOLOGY, 2005, 57 (01) :9-16
[3]   ChIP-chip: considerations for the design, analysis, and application of genome-wide chromatin immunoprecipitation experiments [J].
Buck, MJ ;
Lieb, JD .
GENOMICS, 2004, 83 (03) :349-360
[4]   Transcription activation by catabolite activator protein (CAP) [J].
Busby, S ;
Ebright, RH .
JOURNAL OF MOLECULAR BIOLOGY, 1999, 293 (02) :199-213
[5]   The distribution of RNA polymerase in Escherichia coli is dynamic and sensitive to environmental cues [J].
Cabrera, JE ;
Jin, DJ .
MOLECULAR MICROBIOLOGY, 2003, 50 (05) :1493-1505
[6]   One-step inactivation of chromosomal genes in Escherichia coli K-12 using PCR products [J].
Datsenko, KA ;
Wanner, BL .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2000, 97 (12) :6640-6645
[7]   Organization of supercoil domains and their reorganization by transcription [J].
Deng, S ;
Stein, RA ;
Higgins, NP .
MOLECULAR MICROBIOLOGY, 2005, 57 (06) :1511-1521
[8]   Does RNA polymerase help drive chromosome segregation in bacteria? [J].
Dworkin, J ;
Losick, R .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2002, 99 (22) :14089-14094
[9]   RNA polymerase: Structural similarities between bacterial RNA polymerase and eukaryotic RNA polymerase II [J].
Ebright, RH .
JOURNAL OF MOLECULAR BIOLOGY, 2000, 304 (05) :687-698
[10]   Transcriptome analysis of Crp-dependent catabolite control of gene expression in Escherichia coli [J].
Gosset, G ;
Zhang, ZG ;
Nayyar, SN ;
Cuevas, WA ;
Saier, MH .
JOURNAL OF BACTERIOLOGY, 2004, 186 (11) :3516-3524