Transactivation domain of p53 regulates DNA repair and integrity in human iPS cells

American Journal of Physiology. Heart and Circulatory Physiology
Ramaswamy KannappanJianyi Zhang

Abstract

The role of p53 transactivation domain (p53-TAD), a multifunctional and dynamic domain, on DNA repair and retaining DNA integrity in human induced pluripotent stem cells (hiPSCs) has never been studied. p53-TAD was knocked out in iPSCs using CRISPR/Cas9 and was confirmed by DNA sequencing. p53-TAD knockout (KO) cells were characterized by accelerated proliferation, decreased population doubling time, and unaltered Bcl-2, Bcl-2-binding component 3, insulin-like growth factor 1 receptor, and Bax and altered Mdm2, p21, and p53-induced death domain transcript expression. In p53-TAD KO cells, the p53-regulated DNA repair proteins xeroderma pigmentosum group A, DNA polymerase H, and DNA-binding protein 2 expression were found to be reduced compared with p53 wild-type cells. Exposure to a low dose of doxorubicin (Doxo) induced similar DNA damage and DNA damage response (DDR) as measured by RAD50 and MRE11 expression, checkpoint kinase 2 activation, and γH2A.X recruitment at DNA strand breaks in both cell groups, indicating that silence of p53-TAD does not affect the DDR mechanism upstream of p53. After removal of Doxo, p53 wild-type hiPSCs underwent DNA repair, corrected their damaged DNA, and restored DNA integrity. Conversely, p53-T...Continue Reading

References

Jan 1, 1992·Advances in Experimental Medicine and Biology·M FennemaN S Faithfull
Dec 1, 1995·Current Opinion in Cell Biology·G I EvanE Harrington
Apr 16, 1998·The Journal of Biological Chemistry·E P RogakouW M Bonner
May 28, 1998·The Journal of Biological Chemistry·J ZhuX Chen
Sep 13, 2000·The Journal of Biological Chemistry·J ZhuX Chen
Dec 13, 2000·Molecular and Cellular Biology·O K Mirzoeva, J H Petrini
May 19, 2001·European Journal of Biochemistry·K ItahanaJ Campisi
Jun 23, 2001·Oncogene·P Chène
Feb 13, 2002·Current Opinion in Structural Biology·Karl Peter HopfnerJohn A Tainer
May 4, 2002·Nature Reviews. Molecular Cell Biology·Damien D'Amours, Stephen P Jackson
May 23, 2002·Carcinogenesis·Stephen P Jackson
Sep 23, 2003·Journal of Molecular Biology·Roger DawsonJohannes Buchner
Oct 25, 2003·Oncogene·Wafik S El-Deiry
Dec 3, 2003·Mutation Research·Glaucia R MartinezPaolo Di Mascio
Mar 1, 2005·Cell·David B LombardFrederick W Alt
Jan 25, 2006·Chemico-biological Interactions·M ValkoM Mazur
Mar 18, 2006·Cell Death and Differentiation·K L Harms, X Chen
Aug 9, 2006·Experimental Cell Research·Zhenkun Lou, Junjie Chen
Oct 5, 2007·Nucleic Acids Research·Jian-Hua ChenSusan E Ozanne
Apr 24, 2008·Nature Reviews. Molecular Cell Biology·Todd RileyArnold Levine
Jul 6, 2011·Philosophical Transactions of the Royal Society of London. Series B, Biological Sciences·Keisuke Okita, Shinya Yamanaka
Apr 20, 2012·The Journal of Biological Chemistry·Pierre-Jacques HamardJames J Manfredi
Jan 16, 2013·Current Gene Therapy·Qiang BaiJohn De Vos
Nov 19, 2014·Journal of Molecular Cell Biology·Laura ZanniniGiacomo Buscemi
Nov 1, 2016·Nature·Yuji NakadaHesham A Sadek
Nov 20, 2016·Cold Spring Harbor Perspectives in Medicine·Nitin Raj, Laura D Attardi
Jun 1, 2017·International Journal of Oncology·Xiaofei ZhangGong Yang

❮ Previous
Next ❯

Citations

Feb 15, 2020·American Journal of Physiology. Heart and Circulatory Physiology·Jessica M MillerRamaswamy Kannappan
Jan 3, 2020·Cardiovascular Toxicology·Yi YanDingguo Zhang

❮ Previous
Next ❯

Methods Mentioned

BETA
NMR
Assay
fluorescence microscopy
PCR
FACS
transgenic

Software Mentioned

FlowJo
Cell Cycle
SigmaStat
ImageJ

Related Concepts

Related Feeds

CRISPR (general)

Clustered regularly interspaced short palindromic repeats (CRISPR) are DNA sequences in the genome that are recognized and cleaved by CRISPR-associated proteins (Cas). CRISPR-Cas system enables the editing of genes to create or correct mutations. Discover the latest research on CRISPR here.

CRISPR for Genome Editing

Genome editing technologies enable the editing of genes to create or correct mutations. Clustered regularly interspaced short palindromic repeats (CRISPR) are DNA sequences in the genome that are recognized and cleaved by CRISPR-associated proteins (Cas). Here is the latest research on the use of CRISPR-Cas system in gene editing.

CRISPR Ribonucleases Deactivation

CRISPR-Cas system enables the editing of genes to create or correct mutations. This feed focuses on mechanisms that underlie deactivation of CRISPR ribonucleases. Here is the latest research.

BCL-2 Family Proteins

BLC-2 family proteins are a group that share the same homologous BH domain. They play many different roles including pro-survival signals, mitochondria-mediated apoptosis and removal or damaged cells. They are often regulated by phosphorylation, affecting their catalytic activity. Here is the latest research on BCL-2 family proteins.