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Human histone acetyltransferase 1 (Hat1) acetylates lysine 5 of histone H2A in vivo

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Abstract

The primary structure of Histone Acetyltransferase 1 (Hat1) has been conserved throughout evolution; however, despite its ubiquity, its cellular function is not well characterized. To study its in vivo acetylation pattern and function, we utilized shRNAmir against Hat1 expressed in the well-substantiated HeLa (human cervical cancer) cell line. To reduce the interference by enzymes with similar HAT specificity, we used HeLa cells expressing histone acetyltransferase Tip60 with mutated acetyl-CoA binding site that abrogates its enzyme activity (mutant HeLa-tip60). Two shRNAmir were identified that reduced the expression of the cytoplasmic and nuclear forms of Hat1. Cytosolic protein preparations from these two clones showed decreased levels of acetylation of lysine 5 (K5) and K12 on histone H4, with the concomitant loss of the acetylation of histone H2A at K5. This pattern of decreased acetylation of H2AK5 was well defined in preparations of histone protein and insoluble nuclear-protein (INP) fractions as well. Abrogating the Hat1 expression caused a 74 % decrease in colony-forming efficiency of mutant HeLa-tip60 cells, reduced the size of the colonies by 50 %, and decreased the amounts of proteins with molecular weights below 35 kDa in the INP fractions.

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Acknowledgments

We would like to thank Dr. Avril Woodhead for her critical help with the manuscript preparation and Dr. Rolf Sternglanz for the invaluable support, discussions, and comments throughout the years. We would like to thank Dr. David Schlyer, Dr. John Shanklin, and the entire Biosciences Department for the support; and Dr. Tsuyoshi Ikura for providing the HeLa-TIP60 and HeLa-tip60 cell lines. This article has been authored by Brookhaven Science Associates, LLC under contract number DE-AC02-98CH10886 with the U.S. Department of Energy. The United States Government retains and the publisher, by accepting the article for publication, acknowledges that the United States Government retains a non-exclusive, paid-up, irrevocable, world-wide license to publish or reproduce the published form of this manuscript, or allow others to do so, for United States Government purposes. This work was supported by a grant from the National Aeronautics and Space Administration NNJ08HB63I under Department of Energy Prime Contract DE-AC02-98CH10886 with the Brookhaven National Laboratory (to STT).

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The authors declare that there are no conflicts of interest.

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Authors and Affiliations

  1. Department of Oral Biology and Pathology, Stony Brook University, Stony Brook, NY, 11794, USA

    Juliana I. Tafrova

  2. The Genetics Center, Smithtown, NY, 11787, USA

    Juliana I. Tafrova

  3. Department of Biochemistry and Cell Biology, Stony Brook University, Stony Brook, NY, 11794, USA

    Stefan T. Tafrov

  4. Biology Department, Brookhaven National Laboratory, 50 Bell Avenue, Building 463, Upton, NY, 11973-5000, USA

    Stefan T. Tafrov

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  1. Juliana I. Tafrova
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Correspondence to Stefan T. Tafrov.

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Tafrova, J.I., Tafrov, S.T. Human histone acetyltransferase 1 (Hat1) acetylates lysine 5 of histone H2A in vivo. Mol Cell Biochem 392, 259–272 (2014). https://doi.org/10.1007/s11010-014-2036-0

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  • DOI: https://doi.org/10.1007/s11010-014-2036-0

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