Analysis of U1 small nuclear RNA interaction with cyclin H.

TFIIH is a general transcription and repair factor implicated in RNA polymerase II transcription, nucleotide excision repair, and transcription-coupled repair. Genetic defects in TFIIH lead to three distinct inheritable diseases: xeroderma pigmentosa, Cockayne syndrome, and trichothiodystrophy, with...

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Main Authors: O'Gorman, W, Thomas, B, Kwek, K, Furger, A, Akoulitchev, A
Format: Journal article
Language:English
Published: 2005
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author O'Gorman, W
Thomas, B
Kwek, K
Furger, A
Akoulitchev, A
author_facet O'Gorman, W
Thomas, B
Kwek, K
Furger, A
Akoulitchev, A
author_sort O'Gorman, W
collection OXFORD
description TFIIH is a general transcription and repair factor implicated in RNA polymerase II transcription, nucleotide excision repair, and transcription-coupled repair. Genetic defects in TFIIH lead to three distinct inheritable diseases: xeroderma pigmentosa, Cockayne syndrome, and trichothiodystrophy, with xeroderma pigmentosa patients being highly susceptible to skin cancer. Earlier data revealed that the cyclin H subunit of TFIIH associates with U1 small nuclear RNA, a core-splicing component. In addition to its role in RNA processing U1 small nuclear RNA also regulates diverse stages of transcription by RNA polymerase II both in vivo and in vitro, including abortive initiation and re-initiation. Here we identify structural components of U1 and cyclin H implicated in the direct interaction and show how they affect function. Because of unique features of cyclin H we have developed a new methodology for mapping RNA interaction with the full-length cyclin H polypeptide based on electrospray ionization tandem mass spectrometry. We also demonstrate the importance of U1 stem-loops 1 and 2 for the interaction with cyclin H. Functional assays implicate the identified interaction with U1 in regulation of the activity of the cyclin H associated kinase CDK7.
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spelling oxford-uuid:579bfcd1-5788-443b-ac2b-582032b2b3442022-03-26T16:57:44ZAnalysis of U1 small nuclear RNA interaction with cyclin H.Journal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:579bfcd1-5788-443b-ac2b-582032b2b344EnglishSymplectic Elements at Oxford2005O'Gorman, WThomas, BKwek, KFurger, AAkoulitchev, ATFIIH is a general transcription and repair factor implicated in RNA polymerase II transcription, nucleotide excision repair, and transcription-coupled repair. Genetic defects in TFIIH lead to three distinct inheritable diseases: xeroderma pigmentosa, Cockayne syndrome, and trichothiodystrophy, with xeroderma pigmentosa patients being highly susceptible to skin cancer. Earlier data revealed that the cyclin H subunit of TFIIH associates with U1 small nuclear RNA, a core-splicing component. In addition to its role in RNA processing U1 small nuclear RNA also regulates diverse stages of transcription by RNA polymerase II both in vivo and in vitro, including abortive initiation and re-initiation. Here we identify structural components of U1 and cyclin H implicated in the direct interaction and show how they affect function. Because of unique features of cyclin H we have developed a new methodology for mapping RNA interaction with the full-length cyclin H polypeptide based on electrospray ionization tandem mass spectrometry. We also demonstrate the importance of U1 stem-loops 1 and 2 for the interaction with cyclin H. Functional assays implicate the identified interaction with U1 in regulation of the activity of the cyclin H associated kinase CDK7.
spellingShingle O'Gorman, W
Thomas, B
Kwek, K
Furger, A
Akoulitchev, A
Analysis of U1 small nuclear RNA interaction with cyclin H.
title Analysis of U1 small nuclear RNA interaction with cyclin H.
title_full Analysis of U1 small nuclear RNA interaction with cyclin H.
title_fullStr Analysis of U1 small nuclear RNA interaction with cyclin H.
title_full_unstemmed Analysis of U1 small nuclear RNA interaction with cyclin H.
title_short Analysis of U1 small nuclear RNA interaction with cyclin H.
title_sort analysis of u1 small nuclear rna interaction with cyclin h
work_keys_str_mv AT ogormanw analysisofu1smallnuclearrnainteractionwithcyclinh
AT thomasb analysisofu1smallnuclearrnainteractionwithcyclinh
AT kwekk analysisofu1smallnuclearrnainteractionwithcyclinh
AT furgera analysisofu1smallnuclearrnainteractionwithcyclinh
AT akoulitcheva analysisofu1smallnuclearrnainteractionwithcyclinh