DIRECT EVIDENCE FOR THE EFFECT OF TRANSCRIPTION ON LOCAL DNA SUPERCOILING INVIVO

被引:129
作者
RAHMOUNI, AR
WELLS, RD
机构
[1] UNIV ALABAMA,SCH MED,DEPT BIOCHEM,BIRMINGHAM,AL 35294
[2] UNIV ALABAMA,SCH DENT,BIRMINGHAM,AL 35294
关键词
TRANSCRIPTION; DNA SUPERCOILING; OSMIUM TETROXIDE; Z-DNA;
D O I
10.1016/0022-2836(92)90721-U
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
The B-to-Z structural transition of varying lengths (74 to 14 base-pairs) of (CG) tracts has been used as a superhelicity probe to examine the local topological changes induced by transcription at defined genetic loci in vivo. The local-topology reporter sequences indicate that under steady-state transcription the region upstream from the promoter experiences an increase in negative supercoiling whereas the region downstream from the terminator displays a decrease in negative superhelicity. This result provides direct in vivo evidence for the notion that the translocation of an RNA polymerase elongation complex along the double-helical DNA generates positive supercoils in front of it and negative supercoils behind it. Also, this twin-supercoiled domain model was tested inside a transcribed region where a high degree of negative supercoiling generated by the passage of each individual RNA polymerase was detected. Hence, these data indicate that the induced supercoils are confined to the vicinity of each RNA polymerase complex in a multipolymerase system. © 1992.
引用
收藏
页码:131 / 144
页数:14
相关论文
共 53 条
[1]   VECTORS BEARING A HYBRID TRP-LAC PROMOTER USEFUL FOR REGULATED EXPRESSION OF CLONED GENES IN ESCHERICHIA-COLI [J].
AMANN, E ;
BROSIUS, J ;
PTASHNE, M .
GENE, 1983, 25 (2-3) :167-178
[2]   CHANGES IN THE LINKING NUMBER OF SUPERCOILED DNA ACCOMPANY GROWTH TRANSITIONS IN ESCHERICHIA-COLI [J].
BALKE, VL ;
GRALLA, JD .
JOURNAL OF BACTERIOLOGY, 1987, 169 (10) :4499-4506
[3]   USE OF SITE-SPECIFIC RECOMBINATION AS A PROBE OF DNA-STRUCTURE AND METABOLISM INVIVO [J].
BLISKA, JB ;
COZZARELLI, NR .
JOURNAL OF MOLECULAR BIOLOGY, 1987, 194 (02) :205-218
[4]   NEED FOR DNA TOPOISOMERASE ACTIVITY AS A SWIVEL FOR DNA-REPLICATION FOR TRANSCRIPTION OF RIBOSOMAL-RNA [J].
BRILL, SJ ;
DINARDO, S ;
VOELKELMEIMAN, K ;
STERNGLANZ, R .
NATURE, 1987, 326 (6111) :414-416
[5]   TRANSCRIPTION-DEPENDENT DNA SUPERCOILING IN YEAST DNA TOPOISOMERASE MUTANTS [J].
BRILL, SJ ;
STERNGLANZ, R .
CELL, 1988, 54 (03) :403-411
[6]   GENE ORGANIZATION AND PRIMARY STRUCTURE OF A RIBOSOMAL-RNA OPERON FROM ESCHERICHIA-COLI [J].
BROSIUS, J ;
DULL, TJ ;
SLEETER, DD ;
NOLLER, HF .
JOURNAL OF MOLECULAR BIOLOGY, 1981, 148 (02) :107-127
[7]   PLASMID VECTORS FOR THE SELECTION OF PROMOTERS [J].
BROSIUS, J .
GENE, 1984, 27 (02) :151-160
[8]  
CHOMCZYNSKI P, 1987, ANAL BIOCHEM, V162, P156, DOI 10.1016/0003-2697(87)90021-2
[9]  
DEBOER HA, 1983, P NATL ACAD SCI-BIOL, V80, P21
[10]   DNA SUPERCOILING AND THE ANAEROBIC AND GROWTH-PHASE REGULATION OF TONB GENE-EXPRESSION [J].
DORMAN, CJ ;
BARR, GC ;
BHRIAIN, NN ;
HIGGINS, CF .
JOURNAL OF BACTERIOLOGY, 1988, 170 (06) :2816-2826