Snf2/Swi2-related ATPase Mot1 drives displacement of TATA-binding protein by gripping DNA

Rebekka O. Sprouse, Michael D. Brenowitz, David T. Auble

Research output: Contribution to journalArticle

33 Citations (Scopus)

Abstract

Mot1 is a conserved Snf2/Swi2-related transcriptional regulator that uses ATP hydrolysis to displace TATA-binding protein (TBP) from DNA. Several models of the enzymatic mechanism have been proposed, including Mot1-catalyzed distortion of TBP structure, competition between Mot1 and DNA for the TBP DNA-binding surface, and ATP-driven translocation of Mot1 along DNA. Here, DNase I footprinting studies provide strong support for a 'DNA-based' mechanism of Mot1, which we propose involves ATP-driven DNA translocation. Mot1 forms an asymmetric complex with the TBP core domain (TBPc)-DNA complex, contacting DNA both upstream and within the major groove of the TATA Box. Contact with upstream DNA is required for Mot1-mediated displacement of TBPc from DNA. Using the SsoRad54-DNA complex as a model, DNA-binding residues in Mot1 were identified that are critical for Mot1-TBPc-DNA complex formation and catalytic activity, thus placing Mot1 mechanistically within the helicase superfamily. We also report a novel ATP-independent TBPc displacement activity for Mot1 and describe conformational heterogeneity in the Mot1 ATPase, which is likely a general feature of other enzymes in this class.

Original languageEnglish (US)
Pages (from-to)1492-1504
Number of pages13
JournalEMBO Journal
Volume25
Issue number7
DOIs
StatePublished - Apr 5 2006

Fingerprint

TATA-Box Binding Protein
Adenosine Triphosphatases
DNA
Adenosine Triphosphate
Deoxyribonuclease I
DNA-Binding Proteins
Protein Binding

Keywords

  • ATPase
  • Mot1
  • Snf2
  • Swi2
  • TBP

ASJC Scopus subject areas

  • Genetics
  • Cell Biology

Cite this

Snf2/Swi2-related ATPase Mot1 drives displacement of TATA-binding protein by gripping DNA. / Sprouse, Rebekka O.; Brenowitz, Michael D.; Auble, David T.

In: EMBO Journal, Vol. 25, No. 7, 05.04.2006, p. 1492-1504.

Research output: Contribution to journalArticle

Sprouse, Rebekka O. ; Brenowitz, Michael D. ; Auble, David T. / Snf2/Swi2-related ATPase Mot1 drives displacement of TATA-binding protein by gripping DNA. In: EMBO Journal. 2006 ; Vol. 25, No. 7. pp. 1492-1504.
@article{592cefc8e34f446db22a84b12674e51e,
title = "Snf2/Swi2-related ATPase Mot1 drives displacement of TATA-binding protein by gripping DNA",
abstract = "Mot1 is a conserved Snf2/Swi2-related transcriptional regulator that uses ATP hydrolysis to displace TATA-binding protein (TBP) from DNA. Several models of the enzymatic mechanism have been proposed, including Mot1-catalyzed distortion of TBP structure, competition between Mot1 and DNA for the TBP DNA-binding surface, and ATP-driven translocation of Mot1 along DNA. Here, DNase I footprinting studies provide strong support for a 'DNA-based' mechanism of Mot1, which we propose involves ATP-driven DNA translocation. Mot1 forms an asymmetric complex with the TBP core domain (TBPc)-DNA complex, contacting DNA both upstream and within the major groove of the TATA Box. Contact with upstream DNA is required for Mot1-mediated displacement of TBPc from DNA. Using the SsoRad54-DNA complex as a model, DNA-binding residues in Mot1 were identified that are critical for Mot1-TBPc-DNA complex formation and catalytic activity, thus placing Mot1 mechanistically within the helicase superfamily. We also report a novel ATP-independent TBPc displacement activity for Mot1 and describe conformational heterogeneity in the Mot1 ATPase, which is likely a general feature of other enzymes in this class.",
keywords = "ATPase, Mot1, Snf2, Swi2, TBP",
author = "Sprouse, {Rebekka O.} and Brenowitz, {Michael D.} and Auble, {David T.}",
year = "2006",
month = "4",
day = "5",
doi = "10.1038/sj.emboj.7601050",
language = "English (US)",
volume = "25",
pages = "1492--1504",
journal = "EMBO Journal",
issn = "0261-4189",
publisher = "Nature Publishing Group",
number = "7",

}

TY - JOUR

T1 - Snf2/Swi2-related ATPase Mot1 drives displacement of TATA-binding protein by gripping DNA

AU - Sprouse, Rebekka O.

AU - Brenowitz, Michael D.

AU - Auble, David T.

PY - 2006/4/5

Y1 - 2006/4/5

N2 - Mot1 is a conserved Snf2/Swi2-related transcriptional regulator that uses ATP hydrolysis to displace TATA-binding protein (TBP) from DNA. Several models of the enzymatic mechanism have been proposed, including Mot1-catalyzed distortion of TBP structure, competition between Mot1 and DNA for the TBP DNA-binding surface, and ATP-driven translocation of Mot1 along DNA. Here, DNase I footprinting studies provide strong support for a 'DNA-based' mechanism of Mot1, which we propose involves ATP-driven DNA translocation. Mot1 forms an asymmetric complex with the TBP core domain (TBPc)-DNA complex, contacting DNA both upstream and within the major groove of the TATA Box. Contact with upstream DNA is required for Mot1-mediated displacement of TBPc from DNA. Using the SsoRad54-DNA complex as a model, DNA-binding residues in Mot1 were identified that are critical for Mot1-TBPc-DNA complex formation and catalytic activity, thus placing Mot1 mechanistically within the helicase superfamily. We also report a novel ATP-independent TBPc displacement activity for Mot1 and describe conformational heterogeneity in the Mot1 ATPase, which is likely a general feature of other enzymes in this class.

AB - Mot1 is a conserved Snf2/Swi2-related transcriptional regulator that uses ATP hydrolysis to displace TATA-binding protein (TBP) from DNA. Several models of the enzymatic mechanism have been proposed, including Mot1-catalyzed distortion of TBP structure, competition between Mot1 and DNA for the TBP DNA-binding surface, and ATP-driven translocation of Mot1 along DNA. Here, DNase I footprinting studies provide strong support for a 'DNA-based' mechanism of Mot1, which we propose involves ATP-driven DNA translocation. Mot1 forms an asymmetric complex with the TBP core domain (TBPc)-DNA complex, contacting DNA both upstream and within the major groove of the TATA Box. Contact with upstream DNA is required for Mot1-mediated displacement of TBPc from DNA. Using the SsoRad54-DNA complex as a model, DNA-binding residues in Mot1 were identified that are critical for Mot1-TBPc-DNA complex formation and catalytic activity, thus placing Mot1 mechanistically within the helicase superfamily. We also report a novel ATP-independent TBPc displacement activity for Mot1 and describe conformational heterogeneity in the Mot1 ATPase, which is likely a general feature of other enzymes in this class.

KW - ATPase

KW - Mot1

KW - Snf2

KW - Swi2

KW - TBP

UR - http://www.scopus.com/inward/record.url?scp=33645739547&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33645739547&partnerID=8YFLogxK

U2 - 10.1038/sj.emboj.7601050

DO - 10.1038/sj.emboj.7601050

M3 - Article

VL - 25

SP - 1492

EP - 1504

JO - EMBO Journal

JF - EMBO Journal

SN - 0261-4189

IS - 7

ER -