Abstract
NKAP is a multifunctional nuclear protein that associates with the histone deacetylase HDAC3. Although both NKAP and HDAC3 are critical for hematopoietic stem cell (HSC) maintenance and survival, it was not known whether these two proteins work together. To assess the importance of their association in vivo, serial truncation and alanine scanning was performed on NKAP to identify the minimal binding site for HDAC3. Mutation of either Y352 or F347 to alanine abrogated the association of NKAP with HDAC3, but did not alter NKAP localization or expression. Using a linked conditional deletion/re-expression system in vivo, we demonstrated that re-expression of the Y352A NKAP mutant failed to restore HSC maintenance and survival in mice when endogenous NKAP expression was eliminated using Mx1-cre and poly-IC, whereas re-expression of wild type NKAP maintained the HSC pool. However, Y352A NKAP did restore proliferation in murine embryonic fibroblasts when endogenous NKAP expression was eliminated using ER-cre and tamoxifen. Therefore, Y352 in NKAP is critical for association with HDAC3 and for HSC maintenance and survival but is not important for proliferation of murine embryonic fibroblasts, demonstrating that NKAP functions in differ...Continue Reading
References
Feb 16, 2002·Methods : a Companion to Methods in Enzymology·K J Livak, T D Schmittgen
Mar 9, 2002·Molecular and Cellular Biology·William C HahnRobert A Weinberg
Jan 10, 2006·Genesis : the Journal of Genetics and Development·Caroline BeardRudolf Jaenisch
Mar 22, 2006·Pediatric Research·Michael A Teitell, Hanna K A Mikkola
Dec 7, 2007·Neurobiology of Disease·Lijun WangRaymond P Roos
Feb 22, 2008·The Journal of Biological Chemistry·Katie L KathreinSusan Winandy
May 5, 2009·Immunity·Anthony G PajerowskiVirginia Smith Shapiro
Jul 9, 2010·Blood·Anthony G PajerowskiVirginia Smith Shapiro
Jun 1, 2011·The Journal of Experimental Medicine·Fan-Chi HsuVirginia Smith Shapiro
Mar 14, 2013·Nature Communications·Puspa ThapaVirginia Smith Shapiro
Aug 8, 2013·The Journal of Clinical Investigation·Alyssa R SummersScott W Hiebert
Dec 20, 2013·Nucleic Acids Research·Bhagyashri D BurguteAngelika A Noegel
Apr 30, 2014·Nature Cell Biology·Hyun-Ja Nam, Jan M van Deursen
Jul 3, 2015·Thrombosis and Haemostasis·Leonardo RivadeneyraMirta Schattner
Jul 15, 2015·The Journal of Immunology : Official Journal of the American Association of Immunologists·Fan-Chi HsuVirginia Smith Shapiro
May 18, 2016·The Journal of Immunology : Official Journal of the American Association of Immunologists·Puspa ThapaVirginia Smith Shapiro
Sep 25, 2016·Blood·David KavanaghDavid P J Hunt
Oct 4, 2016·Nature Communications·Teng LiQing Xia
Dec 21, 2016·PloS One·Bhagyashri D BurguteAngelika A Noegel
Jan 26, 2018·Journal of Autoimmunity·Barsha DashVirginia Smith Shapiro