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Quantitation of the DNA tethering effect in long-range DNA looping in vivo and in vitro using the Lac and λ repressors

Efficient and specific interactions between proteins bound to the same DNA molecule can be dependent on the length of the DNA tether that connects them. Measurement of the strength of this DNA tethering effect has been largely confined to short separations between sites, and it is not clear how it c... Full description

Journal Title: Proceedings of the National Academy of Sciences of the United States of America 07 January 2014, Vol.111(1), pp.349-54
Main Author: Priest, David G
Other Authors: Cui, Lun , Kumar, Sandip , Dunlap, David D , Dodd, Ian B , Shearwin, Keith E
Format: Electronic Article Electronic Article
Language: English
Subjects:
Tpm
ID: E-ISSN: 1091-6490 ; PMID: 24344307 Version:1 ; DOI: 10.1073/pnas.1317817111
Link: http://pubmed.gov/24344307
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recordid: medline24344307
title: Quantitation of the DNA tethering effect in long-range DNA looping in vivo and in vitro using the Lac and λ repressors
format: Article
creator:
  • Priest, David G
  • Cui, Lun
  • Kumar, Sandip
  • Dunlap, David D
  • Dodd, Ian B
  • Shearwin, Keith E
subjects:
  • Tpm
  • J Factor
  • Promoter-Enhancer
  • DNA, Bacterial -- Genetics
  • Escherichia Coli -- Genetics
  • Lac Repressors -- Genetics
  • Repressor Proteins -- Genetics
  • Viral Regulatory and Accessory Proteins -- Genetics
ispartof: Proceedings of the National Academy of Sciences of the United States of America, 07 January 2014, Vol.111(1), pp.349-54
description: Efficient and specific interactions between proteins bound to the same DNA molecule can be dependent on the length of the DNA tether that connects them. Measurement of the strength of this DNA tethering effect has been largely confined to short separations between sites, and it is not clear how it contributes to long-range DNA looping interactions, such as occur over separations of tens to hundreds of kilobase pairs in vivo. Here, gene regulation experiments using the LacI and λ CI repressors, combined with mathematical modeling, were used to quantitate DNA tethering inside Escherichia coli cells over the 250- to 10,000-bp range. Although LacI and CI loop DNA in distinct ways, measurements of the tethering effect were very similar for both proteins. Tethering strength decreased with increasing separation, but even at 5- to 10-kb distances, was able to increase contact probability 10- to 20-fold and drive efficient looping. Tethering in vitro with the Lac repressor was measured for the same...
language: eng
source:
identifier: E-ISSN: 1091-6490 ; PMID: 24344307 Version:1 ; DOI: 10.1073/pnas.1317817111
fulltext: fulltext
issn:
  • 10916490
  • 1091-6490
url: Link


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titleQuantitation of the DNA tethering effect in long-range DNA looping in vivo and in vitro using the Lac and λ repressors
creatorPriest, David G ; Cui, Lun ; Kumar, Sandip ; Dunlap, David D ; Dodd, Ian B ; Shearwin, Keith E
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subjectTpm ; J Factor ; Promoter-Enhancer ; DNA, Bacterial -- Genetics ; Escherichia Coli -- Genetics ; Lac Repressors -- Genetics ; Repressor Proteins -- Genetics ; Viral Regulatory and Accessory Proteins -- Genetics
descriptionEfficient and specific interactions between proteins bound to the same DNA molecule can be dependent on the length of the DNA tether that connects them. Measurement of the strength of this DNA tethering effect has been largely confined to short separations between sites, and it is not clear how it contributes to long-range DNA looping interactions, such as occur over separations of tens to hundreds of kilobase pairs in vivo. Here, gene regulation experiments using the LacI and λ CI repressors, combined with mathematical modeling, were used to quantitate DNA tethering inside Escherichia coli cells over the 250- to 10,000-bp range. Although LacI and CI loop DNA in distinct ways, measurements of the tethering effect were very similar for both proteins. Tethering strength decreased with increasing separation, but even at 5- to 10-kb distances, was able to increase contact probability 10- to 20-fold and drive efficient looping. Tethering in vitro with the Lac repressor was measured for the same...
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descriptionEfficient and specific interactions between proteins bound to the same DNA molecule can be dependent on the length of the DNA tether that connects them. Measurement of the strength of this DNA tethering effect has been largely confined to short separations between sites, and it is not clear how it contributes to long-range DNA looping interactions, such as occur over separations of tens to hundreds of kilobase pairs in vivo. Here, gene regulation experiments using the LacI and λ CI repressors, combined with mathematical modeling, were used to quantitate DNA tethering inside Escherichia coli cells over the 250- to 10,000-bp range. Although LacI and CI loop DNA in distinct ways, measurements of the tethering effect were very similar for both proteins. Tethering strength decreased with increasing separation, but even at 5- to 10-kb distances, was able to increase contact probability 10- to 20-fold and drive efficient looping. Tethering in vitro with the Lac repressor was measured for the same...
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abstractEfficient and specific interactions between proteins bound to the same DNA molecule can be dependent on the length of the DNA tether that connects them. Measurement of the strength of this DNA tethering effect has been largely confined to short separations between sites, and it is not clear how it contributes to long-range DNA looping interactions, such as occur over separations of tens to hundreds of kilobase pairs in vivo. Here, gene regulation experiments using the LacI and λ CI repressors, combined with mathematical modeling, were used to quantitate DNA tethering inside Escherichia coli cells over the 250- to 10,000-bp range. Although LacI and CI loop DNA in distinct ways, measurements of the tethering effect were very similar for both proteins. Tethering strength decreased with increasing separation, but even at 5- to 10-kb distances, was able to increase contact probability 10- to 20-fold and drive efficient looping. Tethering in vitro with the Lac repressor was measured for the same...
doi10.1073/pnas.1317817111
pmid24344307
date2014-01-07