In our efforts to understand how transcription may be regulated in Entamoeba histolytica, we have examined if this parasite has conserved enzymatic mechanisms for targeted acetylation and deacetylation of histories. Western blotting indicated that basic nuclear proteins in the size range of 16-23 kDa were acetylated in amebic trophozoites, suggesting histone acetylation. Single representatives of the GNAT and MYST family of histone acetyltransferases (HATs) were identified in the E. histolytica genome and their expression in amebic trophozoites was detected by reverse transcription of RNA followed by the polymerase chain reaction (RT-PCR). Full-length recombinant EhMYST protein demonstrated HAT activity with calf thymus histories and showed a preference for histone H4, similar to the yeast MYST protein, Esa1. However, ehMYST did not complement a yeast esa1 mutation. Histone deacetylase (HDAC) activity was detected in nuclear extracts from E. histolytica, and characteristically, was inhibited by trichostatin A (TSA). Consistent with the observation of HDAC activity, RT-PCR analysis demonstrated that an amebic hdac1 homolog (ehHDAC) is expressed and appropriately spliced in E. histolytica trophozoites. Our results suggest that mechanisms for histone acetylation and deacetylation are operational in E. histolytica. (C) 2004 Elsevier B.V. All rights reserved.
机构:
Penn State Univ, Howard Hughes Med Inst, Dept Biochem & Mol Biol, University Pk, PA 16803 USAPenn State Univ, Howard Hughes Med Inst, Dept Biochem & Mol Biol, University Pk, PA 16803 USA
Carrozza, MJ
;
Utley, RT
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机构:Penn State Univ, Howard Hughes Med Inst, Dept Biochem & Mol Biol, University Pk, PA 16803 USA
Utley, RT
;
Workman, JL
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机构:Penn State Univ, Howard Hughes Med Inst, Dept Biochem & Mol Biol, University Pk, PA 16803 USA
Workman, JL
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Côté, J
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机构:Penn State Univ, Howard Hughes Med Inst, Dept Biochem & Mol Biol, University Pk, PA 16803 USA
机构:
Penn State Univ, Howard Hughes Med Inst, Dept Biochem & Mol Biol, University Pk, PA 16803 USAPenn State Univ, Howard Hughes Med Inst, Dept Biochem & Mol Biol, University Pk, PA 16803 USA
Carrozza, MJ
;
Utley, RT
论文数: 0引用数: 0
h-index: 0
机构:Penn State Univ, Howard Hughes Med Inst, Dept Biochem & Mol Biol, University Pk, PA 16803 USA
Utley, RT
;
Workman, JL
论文数: 0引用数: 0
h-index: 0
机构:Penn State Univ, Howard Hughes Med Inst, Dept Biochem & Mol Biol, University Pk, PA 16803 USA
Workman, JL
;
Côté, J
论文数: 0引用数: 0
h-index: 0
机构:Penn State Univ, Howard Hughes Med Inst, Dept Biochem & Mol Biol, University Pk, PA 16803 USA