Open AccessαTAT1 downregulation induces mitotic catastrophe in HeLa and A549 cells
Author(s) -
Chien Jy,
Tsen Sd,
Chih Cheng Chien,
Liu Hw,
Tung Cy,
Chang-Ping Lin
Publication year - 2016
Publication title -
cell death discovery
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.556
H-Index - 28
ISSN - 2058-7716
DOI - 10.1038/cddiscovery.2016.6
Subject(s) - downregulation and upregulation , microbiology and biotechnology , hela , mitosis , microtubule , a549 cell , caenorhabditis elegans , biology , chemistry , acetylation , cell culture , biochemistry , genetics , gene
α -Tubulin acetyltransferase 1 ( α TAT1) controls reversible acetylation on Lys40 of α -tubulin and modulates multiple cellular functions. α TAT1 depletion induced morphological defects of touch receptor neurons in Caenorhabditis elegans and impaired cell adhesion and contact inhibition in mouse embryonic fibroblasts, however, no morphological or proliferation defects in human RPE-hTERT cells were found after α TAT1-specific siRNA treatment. Here, we compared the effect of three α TAT1-specific shRNAs on proliferation and morphology in two human cell lines, HeLa and A549. The more efficient two shRNAs induced mitotic catastrophe in both cell lines and the most efficient one also decreased F-actin and focal adhesions. Further analysis revealed that α TAT1 downregulation increased γ -H2AX, but not other DNA damage markers p-CHK1 and p-CHK2, along with marginal change in microtubule outgrowth speed and inter-kinetochore distance. Overexpression of α TAT1 could not precisely mimic the distribution and concentration of endogenous acetylated α -tubulin (Ac-Tu), although no overt phenotype change was observed, meanwhile, this could not completely prevent α TAT1 downregulation-induced deficiencies. We therefore conclude that efficient α TAT1 downregulation could impair actin architecture and induce mitotic catastrophe in HeLa and A549 cells through mechanisms partly independent of Ac-Tu.