Hoffmann, R. A wiki for the life sciences where authorship matters. Nature Genetics (2008)

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Sukhodolets, K.E. et al.

Sukhodolets, Hickman, Agarwal, Sukhodolets, Obungu, Novotny, Crabtree, Chandrasekharappa, Collins, Spiegel, Burns, Marx,

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

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.[1]

References

The 32-kilodalton subunit of replication protein A interacts with menin, the product of the MEN1 tumor suppressor gene. Sukhodolets, K.E., Hickman, A.B., Agarwal, S.K., Sukhodolets, M.V., Obungu, V.H., Novotny, E.A., Crabtree, J.S., Chandrasekharappa, S.C., Collins, F.S., Spiegel, A.M., Burns, A.L., Marx, S.J. Mol. Cell. Biol. (2003) [Pubmed]

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