RBT1, a novel transcriptional co-activator, binds the second subunit of Replication Protein A

John M Cho; Daniel J Song; Josee Bergeron; Naciba Benlimame; Marc S Wold; Moulay A Alaoui-Jamali

doi:10.1093/nar/28.18.3478

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RBT1, a novel transcriptional co-activator, binds the second subunit of Replication Protein A

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RBT1, a novel transcriptional co-activator, binds the second subunit of Replication Protein A

John M Cho, Daniel J Song, Josee Bergeron, Naciba Benlimame, Marc S Wold and Moulay A Alaoui-Jamali

Nucleic acids research, Vol.28(18), pp.3478-3485

09/15/2000

DOI: 10.1093/nar/28.18.3478

PMCID: PMC110737

PMID: 10982866

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Abstract

Replication Protein A (RPA) is required for DNA recombination, repair and replication in all eukaryotes. RPA participation in these pathways is mediated by single-stranded DNA binding and protein interactions. We herein identify a novel protein, Replication Protein Binding Trans-Activator (RBT1), in a yeast two-hybrid assay employing the second subunit of human RPA (RPA32) as bait. RBT1–RPA32 binding was confirmed by glutathione S -transferase pull-down and co-immunoprecipitation. Fluorescence microscopy indicates that green fluorescence protein-tagged RBT1 is localized to the nucleus in vivo . RBT1 mRNA expression, determined by semi-quantitative RT–PCR, is significantly higher in cancer cell lines MCF-7, ZR-75, SaOS-2 and H661, compared to the cell lines normal non-immortalized human mammary epithelial cells and normal non-immortalized human bronchial epithelial cells. Further, yeast and mammalian one-hybrid analysis shows that RBT1 is a strong transcriptional co-activator. Interestingly, mammalian transactivation data is indicative of significant variance between cell lines; the GAL4–RBT1 fusion protein has significantly higher transcriptional activity in human cancer cells compared to human normal primary non-immortalized epithelial cells. We propose that RBT1 is a novel transcriptional co-activator that interacts with RPA, and has significantly higher activity in transformed cells.

Details

Title: Subtitle: RBT1, a novel transcriptional co-activator, binds the second subunit of Replication Protein A
Creators: John M Cho - Departments of Medicine, Oncology and Pharmacology, Lady Davis Institute for Medical Research of the Jewish General Hospital, McGill University, Montreal, QC H3T 1E2, Canada and
Daniel J Song - Departments of Medicine, Oncology and Pharmacology, Lady Davis Institute for Medical Research of the Jewish General Hospital, McGill University, Montreal, QC H3T 1E2, Canada and
Josee Bergeron - Departments of Medicine, Oncology and Pharmacology, Lady Davis Institute for Medical Research of the Jewish General Hospital, McGill University, Montreal, QC H3T 1E2, Canada and
Naciba Benlimame - Departments of Medicine, Oncology and Pharmacology, Lady Davis Institute for Medical Research of the Jewish General Hospital, McGill University, Montreal, QC H3T 1E2, Canada and
Marc S Wold - Departments of Medicine, Oncology and Pharmacology, Lady Davis Institute for Medical Research of the Jewish General Hospital, McGill University, Montreal, QC H3T 1E2, Canada and
Moulay A Alaoui-Jamali - Departments of Medicine, Oncology and Pharmacology, Lady Davis Institute for Medical Research of the Jewish General Hospital, McGill University, Montreal, QC H3T 1E2, Canada and
Resource Type: Journal article
Publication Details: Nucleic acids research, Vol.28(18), pp.3478-3485
DOI: 10.1093/nar/28.18.3478
PMID: 10982866
PMCID: PMC110737
NLM abbreviation: Nucleic Acids Res
ISSN: 0305-1048
eISSN: 1362-4962
Publisher: Oxford University Press; Oxford, UK
Language: English
Date published: 09/15/2000
Academic Unit: Radiation Oncology; Biochemistry and Molecular Biology
Record Identifier: 9984025293502771

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