Identification of delta helicase as the bovine homolog of HUPF1: demonstration of an interaction with the third subunit of DNA polymerase delta

L Michael Carastro, Cheng-Keat Tan, Manuel Selg, Hans-Martin Jack, Antero G So, Kathleen M Downey

Publikation: Beitrag in FachzeitschriftArtikelBegutachtung

27 Zitate (Scopus)

Abstract

Delta helicase is a 5′ to 3′ DNA helicase that partially co-purifies with DNA polymerase delta (pol delta) from fetal bovine thymus tissue. We describe the resolution of delta helicase from pol delta on heparin-agarose chromatography and its purification to apparent homogeneity by affinity purification on single-stranded DNA-cellulose chromatography, unique-sequence RNA-agarose chromatography, and ceramic hydroxyapatite chromatography. Delta helicase isolated from fetal bovine thymus had an apparent M r of 115 kDa in SDS-PAGE, and photocrosslinked to [α- 32P]ATP. Tandem mass spectrometry peptide mass data derived from the bovine polypeptide matched to human UPF1 (HUPF1), a 5′ to 3′ RNA and DNA helicase, and a requisite component of the mRNA surveillance complex. Antisera against HUPF1 cross-reacted with delta helicase on western analysis, and delta helicase activity was immuno-inactivated by pre-incubation with antibodies to HUPF1, suggesting that delta helicase is the bovine homolog of HUPF1. Immunoprecipitation experiments demonstrated that HUPF1 interacts with the 66-kDa third subunit of pol delta in vivo.

OriginalspracheEnglisch
Seiten (von - bis)2232-2243
Seitenumfang12
FachzeitschriftNucleic Acids Research
Jahrgang30
Ausgabenummer10
DOIs
PublikationsstatusVeröffentlicht - 10 Mai 2002

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