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PMID: 8631670 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Surveying a supercoil domain by using the gamma delta resolution system in Salmonella typhimurium.

Journal of bacteriology ·Vol. 178 ·No. 10 ·1996-05-00 ·Pages 2825-35

Higgins NP, Yang X, Fu Q, Roth JR

Abstract

A genetic system was developed to investigate the supercoil structure of bacterial chromosomes. New res-carrying transposons were derived from MudI1734 (MudJr1 and MudJr2) and Tn10 (Tn10dGn). The MudJr1 and MudJr2 elements each have a res site in opposite orientation so that when paired with a Tn10dGn element in the same chromosome, one MudJr res site will be ordered as a direct repeat. Deletion formation was studied in a nonessential region (approximately 100 kb) that extends from the his operon through the cob operon. Strains with a MudJr insertion in the cobT gene at the 5' end of the cob operon plus a Tn10dGn insertion positioned either clockwise or counterclockwise from cobT were exposed to a burst of RES protein. Following a pulse of resolvase expression, deletion formation was monitored by scoring the loss of the Lac+ phenotype or by loss of tetracycline resistance. In exponentially growing populations, deletion products appeared quickly in some cells (in 10 min) but also occurred more than an hour after RES induction. The frequency of deletion (y) diminished with increasing distance (x) between res sites. Results from 15 deletion intervals fit the exponential equation y = 120 . 10(-0.02x). We found that res sites can be plectonemically interwound over long distances ( > 100 kb) and that barriers to supercoil diffusion are placed stochastically within the 43- to 45-min region of the chromosome.

MeSH Terms
Cell Division Chromosomes, Bacterial/ultrastructure DNA Nucleotidyltransferases/metabolism DNA Transposable Elements DNA, Bacterial/ultrastructure DNA, Superhelical/ultrastructure Genetic Vectors Lac Operon Mutagenesis, Insertional Phenotype Salmonella typhimurium/genetics,ultrastructure Sensitivity and Specificity Sequence Deletion Tetracycline Resistance/genetics Transposases
Chemicals
DNA Transposable Elements DNA, Bacterial DNA, Superhelical DNA Nucleotidyltransferases Transposases
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Higgins N P
Department of Biochemistry, University of Alabama at Birmingham, 35294-2170, USA. nphiggins@bmg.bhs.uab.edu
Yang X
Fu Q
Roth J R
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/jb.178.10.2825-2835.1996
Email
nphiggins@bmg.bhs.uab.edu
Published
1996-05-00
Pages
2825-35
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC178017
Subset
IM
Grants
NIGMS NIH HHS · GM33143 · United States
NIGMS NIH HHS · GM34804 · United States
External Links
PubMed source DOI: 10.1128/jb.178.10.2825-2835.1996 PMC Full Text View journal details
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