MYH9 binds to dNTPs via deoxyribose moiety and plays an important role in DNA synthesis
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Pratima Nangia-Makker1,2, Malathy P.V. Shekhar1,2,3, Victor Hogan1, Vitaly Balan4 and Avraham Raz1,2,3
1 Barbara Ann Karmanos Cancer Institute, School of Medicine, Wayne State University, Detroit, MI 48201, USA
2 Department of Oncology, School of Medicine, Wayne State University, Detroit, MI 48201, USA
3 Department of Pathology, School of Medicine, Wayne State University, Detroit, MI 48201, USA
4 Guardant Health, Redwood City, CA 94063, USA
Keywords: myosin II; MYH9; dNTP; deoxyribose-5-phosphate; DNA synthesis
Received: January 25, 2022 Accepted: March 04, 2022 Published: March 14, 2022
The accepted notion of dNTP transport following cytoplasmic biosynthesis is ‘facilitated diffusion’; however, whether this alone is sufficient for moving dNTPs for DNA synthesis remains an open question. The data presented here show that the MYH9 gene encoded heavy chain of non-muscle myosin IIA binds dNTPs potentially serving as a ‘reservoir’. Pull-down assays showed that MYH9 present in the cytoplasmic, mitochondrial and nuclear compartments bind to DNA and this interaction is inhibited by dNTPs and 2-deoxyribose-5-phosphate (dRP) suggesting that MYH9-DNA binding is mediated via pentose sugar recognition. Direct dNTP-MYH9 binding was demonstrated by ELISA and a novel PCR-based method, which showed that all dNTPs bind to MYH9 with varying efficiencies. Cellular thermal shift assays showed that MYH9 thermal stability is enhanced by dNTPs. MYH9 siRNA transfection or treatment with myosin II selective inhibitors ML7 or blebbistatin decreased cell proliferation compared to controls. EdU labeling and cell cycle analysis by flow cytometry confirmed MYH9 siRNA and myosin II inhibitors decreased progression to S-phase with accumulation of cells in G0/G1 phase. Taken together, our data suggest a novel role for MYH9 in dNTP binding and DNA synthesis.
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