Molecular cloning of a novel human gene encoding histone acetyltransferase-like protein involved in transcriptional activation of hTERT.
To isolate proteins involved in hTERT transcriptional regulation, the HeLa cDNA library was screened using the hTERT promoter-based yeast one-hybrid assay. A positive clone was rescued and proved to contain an open reading frame and the upstream coding sequences were obtained by 5'-RACE. The assembled full cDNA consisted of a 2.5 kb reading frame encoding 834 amino acids, in which a conserved N-acetyltransferase domain (GNAT family) was searched out in bioinformatics, and thus named as hALP (human N-acetyltransferase-like protein, GenBank Accession No. AF 489535). The expression of native hALP was identified in HeLa cells and proved to distribute in the cellular nucleus. The binding potential of hALP to hTERT promoter was confirmed by EMSA and the interacting sequence involved to -201- to -56-nt upstream region of the promoter. On transfection assay, hALP could obviously transactivate hTERT promoter and stimulate endogenous telomerase activity of cells. The analysis on histone acetyltransferase showed that hALP could specifically acetylate free histones in vitro. The investigation suggested that hALP influences the activity of histone acetylation and could up-regulate telomerase activity through transactivation of hTERT promoter.
Department of Pathology, Health Science Center of Peking University, 38 Xueyuan road, Haidian district, Beijing 100083, China.
Junjie
J
Lv
Haijing
H
Liu
Qiang
Q
Wang
Zhiwei
Z
Tang
Lin
L
Hou
Bo
B
Zhang
D000595:Amino Acid Sequence; D001215:Asparaginase / Q000235:genetics* / Q000378:metabolism*; D001324:Autoantigens / Q000235:genetics* / Q000378:metabolism*; D003001:Cloning, Molecular / Q000379:methods*; D004268:DNA-Binding Proteins; D004789:Enzyme Activation; D015971:Gene Expression Regulation, Enzymologic / Q000502:physiology*; D006367:HeLa Cells; D006801:Humans; D008969:Molecular Sequence Data; D015202:Protein Engineering / Q000379:methods; D019098:Telomerase / Q000235:genetics* / Q000378:metabolism*; D015533:Transcriptional Activation / Q000502:physiology*
D001324:Autoantigens; D004268:DNA-Binding Proteins; D019098:Telomerase; C466840:ASRGL1 protein, human; D001215:Asparaginase