MutSβ Abundance and Msh3 ATP Hydrolysis Activity Are Important Drivers of CTG•CAG Repeat Expansions
CTG•CAG repeat expansions cause at least twelve inherited neurological diseases. Expansions require the presence, not the absence, of the mismatch repair protein MutSβ (Msh2-Msh3 heterodimer). To evaluate properties of MutSβ that drive expansions, previous studies have tested under-expression, ATPase function or polymorphic variants of Msh2 and Msh3, but in disparate experimental systems. Additionally, some variants destabilize MutSβ, potentially masking the effects of biochemical alterations of the variations. Here, human Msh3 was mutated to selectively inactivate MutSβ. Msh3−/− cells are severely defective for CTG•CAG repeat expansions but show full activity on contractions. Msh3−/− cells provide a single, isogenic system to add back Msh3 and test key biochemical features of MutSβ on expansions. Msh3 overexpression led to high expansion activity and elevated levels of MutSβ complex, indicating that MutSβ abundance drives expansions. An ATPase-defective Msh3 expressed at normal levels was as defective in expansions as Msh3−/− cells, indicating that Msh3 ATPase function is critical for expansions. Expression of two Msh3 polymorphic variants at normal levels showed no detectable change in expansions, suggesting these polymorphisms primarily affect Msh3 protein stability, not activity. In summary, CTG•CAG expansions are limited by the abundance of MutSβ and rely heavily on Msh3 ATPase function.
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Science Foundation Ireland [10/IN.1/B2973 to R.S.L]; Irish Research Council Government of Ireland Postgraduate award [GOIPG/2013/638]; Beckman fund scholarship, National University of Ireland Galway and the Thomas Crawford Hayes fund, National University of Ireland Galway [all to N.K]; National Institutes of Health [R01GM112702 to G.-M.L., T32ES02766 to K.C.]. Funding for open access charge: Irish Research Council.
Supplementary Data are available at NAR Online.
Keogh, Norma; Chan, Kara Y.; Li, Guo-Min; and Lahue, Robert S., "MutSβ Abundance and Msh3 ATP Hydrolysis Activity Are Important Drivers of CTG•CAG Repeat Expansions" (2017). Toxicology and Cancer Biology Faculty Publications. 70.
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Published in Nucleic Acids Research, v. 45, no. 17, p. 10068-10078.
© The Author(s) 2017. Published by Oxford University Press on behalf of Nucleic Acids Research.
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