Arte da memória e arquitetura

Rocca, Renata La

doi:10.11606/d.18.2007.tde-20032008-104738

Cited by 1 publication

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“…Antibodies used for immunoprecipitation were anti-MYC (rabbit polyclonal C19, Santa Cruz, CA, USA), anti-MAX (rabbit polyclonal, La Rocca et al, 1994), anti-MSH2 (rabbit polyclonal Ab3 Calbiochem), anti-MLH1 (rabbit polyclonal Ab2, Calbiochem) or preimmune serum (Pre-IS, taken from the rabbit used to raise the anti-MYC serum). Antibodies used for Western analysis were MSH2 (mouse monoclonal Ab2, Calbiochem) and MLH1 (mouse monoclonal G168-15, Pharminogen).…”

Section: Immunoprecipitation Of Cell Extractsmentioning

confidence: 99%

Interactions of the DNA mismatch repair proteins MLH1 and MSH2 with c-MYC and MAX

et al. 2003

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MSH2 and MLH1 have a central role in correcting mismatches in DNA occurring during DNA replication and have been implicated in the engagement of apoptosis induced by a number of cytotoxic anticancer agents. The function of MLH1 is not clearly defined, although it is required for mismatch repair (MMR) and engagement of apoptosis after certain types of DNA damage. In order to identify other partners of MLH1 that may be involved in signalling MMR or apoptosis, we used human MLH1 in yeast two-hybrid screens of normal human breast and ovarian cDNA libraries. As well as known partners of MLH1 such as PMS1, MLH3 and MBD4, we identified the carboxy terminus of the human c-MYC protooncogene as an interacting sequence. We demonstrate, both in vitro by yeast two-hybrid and GST-fusion pulldown experiments, as well as in vivo by coimmunoprecipitation from human tumour cell extracts, that MLH1 interacts with the c-MYC protein. We further demonstrate that the heterodimeric partner of c-MYC, MAX, interacts with a different MMR protein, MSH2, both in vitro and in vivo. Using an inducible c-MYC-ERt fusion gene, we show that elevated c-MYC expression leads to an increased HGPRT mutation rate of Rat1 cells and an increase in the number of frameshift mutants at the HGPRT locus. The effect on HGPRT mutation rate is small (2-3-fold), but is consistent with deregulated c-MYC expression partially inhibiting MMR activity.

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Section: Immunoprecipitation Of Cell Extractsmentioning

confidence: 99%