The 32-kilodalton subunit of replication protein A interacts with menin, the product of the MEN1 tumor suppressor gene

Karen E. Sukhodolets, Alison B. Hickman, Sunita K. Agarwal, Maxim V. Sukhodolets, Victor H. Obungu, Elizabeth A. Novotny, Judy S. Crabtree, Settara C. Chandrasekharappa, Francis S. Collins, Allen M. Spiegel, A. Lee Burns, Stephen J. Marx

Research output: Contribution to journalArticle

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Abstract

Menin is a 70-kDa protein encoded by MEN1, the tumor suppressor gene disrupted in multiple endocrine neoplasia type 1. In a yeast two-hybrid system based on reconstitution of Ras signaling, menin was found to interact with the 32-kDa subunit (RPA2) of replication protein A (RPA), a heterotrimeric protein required for DNA replication, recombination, and repair. The menin-RPA2 interaction was confirmed in a conventional yeast two-hybrid system and by direct interaction between purified proteins. Menin-RPA2 binding was inhibited by a number of menin missense mutations found in individuals with multiple endocrine neoplasia type 1, and the interacting regions were mapped to the N-terminal portion of menin and amino acids 43 to 171 of RPA2. This region of RPA2 contains a weak single-stranded DNA-binding domain, but menin had no detectable effect on RPA-DNA binding in vitro. Menin bound preferentially in vitro to free RPA2 rather than the RPA heterotrimer or a subcomplex consisting of RPA2 bound to the 14-kDa subunit (RPA3). However, the 70-kDa subunit (RPA1) was coprecipitated from HeLa cell extracts along with RPA2 by menin-specific antibodies, suggesting that menin binds to the RPA heterotrimer or a novel RPA1-RPA2-containing complex in vivo. This finding was consistent with the extensive overlap in the nuclear localization patterns of endogenous menin, RPA2, and RPA1 observed by immunofluorescence.

Original languageEnglish (US)
Pages (from-to)493-509
Number of pages17
JournalMolecular and Cellular Biology
Volume23
Issue number2
DOIs
StatePublished - Jan 2003
Externally publishedYes

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Replication Protein A
Multiple Endocrine Neoplasia Type 1
Tumor Suppressor Genes
Two-Hybrid System Techniques
Proteins
A-Form DNA
Recombinational DNA Repair
Single-Stranded DNA
Missense Mutation
Cell Extracts
DNA Replication
HeLa Cells
DNA Repair
Fluorescent Antibody Technique
Amino Acids
Antibodies

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Cell Biology

Cite this

Sukhodolets, K. E., Hickman, A. B., Agarwal, S. K., Sukhodolets, M. V., Obungu, V. H., Novotny, E. A., ... Marx, S. J. (2003). The 32-kilodalton subunit of replication protein A interacts with menin, the product of the MEN1 tumor suppressor gene. Molecular and Cellular Biology, 23(2), 493-509. https://doi.org/10.1128/MCB.23.2.493-509.2003

The 32-kilodalton subunit of replication protein A interacts with menin, the product of the MEN1 tumor suppressor gene. / Sukhodolets, Karen E.; Hickman, Alison B.; Agarwal, Sunita K.; Sukhodolets, Maxim V.; Obungu, Victor H.; Novotny, Elizabeth A.; Crabtree, Judy S.; Chandrasekharappa, Settara C.; Collins, Francis S.; Spiegel, Allen M.; Burns, A. Lee; Marx, Stephen J.

In: Molecular and Cellular Biology, Vol. 23, No. 2, 01.2003, p. 493-509.

Research output: Contribution to journalArticle

Sukhodolets, KE, Hickman, AB, Agarwal, SK, Sukhodolets, MV, Obungu, VH, Novotny, EA, Crabtree, JS, Chandrasekharappa, SC, Collins, FS, Spiegel, AM, Burns, AL & Marx, SJ 2003, 'The 32-kilodalton subunit of replication protein A interacts with menin, the product of the MEN1 tumor suppressor gene', Molecular and Cellular Biology, vol. 23, no. 2, pp. 493-509. https://doi.org/10.1128/MCB.23.2.493-509.2003
Sukhodolets, Karen E. ; Hickman, Alison B. ; Agarwal, Sunita K. ; Sukhodolets, Maxim V. ; Obungu, Victor H. ; Novotny, Elizabeth A. ; Crabtree, Judy S. ; Chandrasekharappa, Settara C. ; Collins, Francis S. ; Spiegel, Allen M. ; Burns, A. Lee ; Marx, Stephen J. / The 32-kilodalton subunit of replication protein A interacts with menin, the product of the MEN1 tumor suppressor gene. In: Molecular and Cellular Biology. 2003 ; Vol. 23, No. 2. pp. 493-509.
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