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PMID: 7937133 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

The xeroderma pigmentosum group B protein ERCC3 produced in the baculovirus system exhibits DNA helicase activity.

Nucleic acids research ·Vol. 22 ·No. 20 ·1994-10-11 ·Pages 4095-102

Ma L, Siemssen ED, Noteborn HM, van der Eb AJ

Abstract

The XPB/ERCC3 gene corrects the nucleotide excision-repair defect in the human hereditary disease xeroderma pigmentosum group B and encodes the largest subunit of the basal transcription factor BTF2/TFIIH. The primary sequence of the XPB/ERCC3 protein features the hallmarks of seven helicase motifs found in many known and putative helicases or helicase-related proteins. Recently, the multiprotein BTF2/TFIIH complex has been found to be associated with DNA helicase activity. To explore the properties and functions of XPB/ERCC3, we have used the baculovirus/insect-cell expression system to produce recombinant protein. We report here the construction and analysis of recombinant baculovirus expressing XPB/ERCC3. The XPB/ERCC3 protein is synthesized at a relatively high level in baculovirus-infected insect cells. While the majority of XPB/ERCC3 end up in the insoluble fraction of insect cell lysates, a minor fraction of recombinant protein is present in soluble form which can be purified under native conditions. We have found that a DNA helicase activity is associated with the purified XPB/ERCC3 protein, suggesting that XPB/ERCC3 may function as a DNA helicase in local unwinding of DNA template both in the context of transcription and nucleotide excision repair.

MeSH Terms
Amino Acid Sequence Animals Baculoviridae/genetics Base Sequence DNA Helicases/metabolism DNA Repair/genetics DNA-Binding Proteins/chemistry,genetics,metabolism Gene Expression Genetic Vectors Molecular Sequence Data Recombinant Proteins/isolation & purification,metabolism Spodoptera/metabolism Transfection Xeroderma Pigmentosum/genetics
Chemicals
DNA-Binding Proteins Recombinant Proteins XPBC-ERCC-3 protein DNA Helicases
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Ma L
Laboratory for Molecular Carcinogenesis, Sylvius Laboratories, Leiden University, The Netherlands.
Siemssen E D
Noteborn H M
van der Eb A J
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/22.20.4095
Published
1994-10-11
Pages
4095-102
Language
English
Region
England
NLM ID
0411011
PMCID
PMC331895
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/22.20.4095 PMC Full Text View journal details
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