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PMID:11445587

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Citation

Sandrock, B and Egly, JM (2001) A yeast four-hybrid system identifies Cdk-activating kinase as a regulator of the XPD helicase, a subunit of transcription factor IIH. J. Biol. Chem. 276:35328-33

Abstract

To understand the role of the various components of TFIIH, a DNA repair/transcription factor, a yeast four-hybrid system was designed. When the ternary Cdk-activating kinase (CAK) complex composed of Cdk7, cyclin H, and MAT1 was used as bait, the xeroderma pigmentosum (XP) D helicase of transcription factor IIH (TFIIH), among other proteins, was identified as an interacting partner. Deletion mutant analyses demonstrated that the coiled-coil and the hydrophobic domains of MAT1 interlink the CAK complex directly with the N-terminal domain of XPD. Using immunoprecipitates from cells coinfected with baculoviruses, we further validated the bridging function of XPD, which anchors CAK to the core TFIIH. In addition we show that upon interaction with MAT1, CAK inhibits the helicase activity of XPD. This inhibition is overcome upon binding to p44, a subunit of the core TFIIH. It is not surprising that under these conditions some XPD mutations affect interactions not only with p44, but also with MAT1, thus preventing either the CAK inhibitory function within CAK.XPD and/or the role of CAK within TFIIH and, consequently, explaining the variety of the XP phenotypes.

Links

PubMed Online version:10.1074/jbc.M105570200

Keywords

Base Sequence; Cyclin H; Cyclin-Dependent Kinases; Cyclins/metabolism; DNA Helicases/genetics; DNA Helicases/metabolism; DNA Primers; DNA-Binding Proteins; Molecular Probe Techniques; Mutation; Phenotype; Protein-Serine-Threonine Kinases/genetics; Protein-Serine-Threonine Kinases/metabolism; Protein-Serine-Threonine Kinases/physiology; Proteins/genetics; Proteins/metabolism; Transcription Factor TFIIH; Transcription Factors/chemistry; Transcription Factors/metabolism; Transcription Factors, TFII; Xeroderma Pigmentosum Group D Protein

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

HUMAN:ERCC2

part_of

GO:0070516: CAK-ERCC2 complex

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:ERCC2

enables

GO:0043139: 5'-3' DNA helicase activity

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

HUMAN:ERCC2

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:P51948

F

Seeded From UniProt

complete

HUMAN:ERCC2

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:Q13888

F

Seeded From UniProt

complete

HUMAN:ERCC2

enables

GO:0030674: protein-macromolecule adaptor activity

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:P51948,UniProtKB:Q13888

F

Seeded From UniProt

complete

HUMAN:MAT1

part_of

GO:0019907: cyclin-dependent protein kinase activating kinase holoenzyme complex

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:MAT1

involved_in

GO:1905775: negative regulation of DNA helicase activity

ECO:0000314: direct assay evidence used in manual assertion

P

  • has_input:(UniProtKB:P18074)

Seeded From UniProt

complete

HUMAN:MAT1

enables

GO:0047485: protein N-terminus binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:P18074

F

Seeded From UniProt

complete

HUMAN:TF2H2

part_of

GO:0000439: transcription factor TFIIH core complex

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:TF2H2

involved_in

GO:1905776: positive regulation of DNA helicase activity

ECO:0000314: direct assay evidence used in manual assertion

P

  • has_input:(UniProtKB:P18074)

Seeded From UniProt

complete

HUMAN:TF2H2

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:P18074

F

Seeded From UniProt

complete


See also

References

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