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PMID: 2643114 Published · ppublish English Journal Article Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

In vivo DNA loops in araCBAD: size limits and helical repeat.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 86 ·No. 2 ·1989-01-00 ·Pages 476-80

Lee DH, Schleif RF

Abstract

Formation of a DNA loop by AraC proteins bound at the araI and araO2 sites, whose center-to-center distance is 211 base pairs, is necessary for repression of the araBAD promoter, PBAD, of Escherichia coli. To determine the upper and lower size limits of the loop, we constructed PBAD-reporter gene fusion plasmids with various spacings between araI and araO2 and measured their levels of expression. Spacings larger than about 500 base pairs resulted in elimination of detectable repression. No lower limit to spacing was found, suggesting that AraC protein itself possesses significant flexibility and its bending substantially aids formation of small loops. As the spacing between araI and araO2 varied, the activity of PBAD oscillated with an 11.1-base-pair periodicity, implying that the in vivo helical repeat of this DNA is 11.1 base pairs per turn.

MeSH Terms
AraC Transcription Factor Arabinose/genetics Bacterial Proteins DNA, Bacterial/genetics,metabolism Escherichia coli/genetics Escherichia coli Proteins Gene Expression Regulation Mutation Nucleic Acid Conformation Plasmids Promoter Regions, Genetic Repressor Proteins/genetics,metabolism Transcription Factors/genetics
Chemicals
AraC Transcription Factor AraC protein, E coli Bacterial Proteins DNA, Bacterial Escherichia coli Proteins Repressor Proteins Transcription Factors Arabinose
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Lee D H
Biochemistry Department, Brandeis University, Waltham, MA 02254.
Schleif R F
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.86.2.476
Published
1989-01-00
Pages
476-80
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC286493
Subset
IM
External Links
PubMed source DOI: 10.1073/pnas.86.2.476 PMC Full Text View journal details
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