PMID- 21931855
OWN - NLM
STAT- MEDLINE
DCOM- 20120228
LR  - 20240313
IS  - 1932-6203 (Electronic)
IS  - 1932-6203 (Linking)
VI  - 6
IP  - 9
DP  - 2011
TI  - Complex SUMO-1 regulation of cardiac transcription factor Nkx2-5.
PG  - e24812
LID - 10.1371/journal.pone.0024812 [doi]
LID - e24812
AB  - Reversible post-translational protein modifications such as SUMOylation add 
      complexity to cardiac transcriptional regulation. The homeodomain transcription 
      factor Nkx2-5/Csx is essential for heart specification and morphogenesis. It has 
      been previously suggested that SUMOylation of lysine 51 (K51) of Nkx2-5 is 
      essential for its DNA binding and transcriptional activation. Here, we confirm 
      that SUMOylation strongly enhances Nkx2-5 transcriptional activity and that 
      residue K51 of Nkx2-5 is a SUMOylation target. However, in a range of cultured 
      cell lines we find that a point mutation of K51 to arginine (K51R) does not 
      affect Nkx2-5 activity or DNA binding, suggesting the existence of additional 
      Nkx2-5 SUMOylated residues. Using biochemical assays, we demonstrate that Nkx2-5 
      is SUMOylated on at least one additional site, and this is the predominant site 
      in cardiac cells. The second site is either non-canonical or a "shifting" site, 
      as mutation of predicted consensus sites and indeed every individual lysine in 
      the context of the K51R mutation failed to impair Nkx2-5 transcriptional 
      synergism with SUMO, or its nuclear localization and DNA binding. We also observe 
      SUMOylation of Nkx2-5 cofactors, which may be critical to Nkx2-5 regulation. Our 
      data reveal highly complex regulatory mechanisms driven by SUMOylation to 
      modulate Nkx2-5 activity.
FAU - Costa, Mauro W
AU  - Costa MW
AD  - Developmental and Stem Cell Biology Division, Victor Chang Cardiac Research 
      Institute, Darlinghurst, New South Wales, Australia.
FAU - Lee, Stella
AU  - Lee S
FAU - Furtado, Milena B
AU  - Furtado MB
FAU - Xin, Li
AU  - Xin L
FAU - Sparrow, Duncan B
AU  - Sparrow DB
FAU - Martinez, Camila G
AU  - Martinez CG
FAU - Dunwoodie, Sally L
AU  - Dunwoodie SL
FAU - Kurtenbach, Eleonora
AU  - Kurtenbach E
FAU - Mohun, Tim
AU  - Mohun T
FAU - Rosenthal, Nadia
AU  - Rosenthal N
FAU - Harvey, Richard P
AU  - Harvey RP
LA  - eng
GR  - MC_U117562103/MRC_/Medical Research Council/United Kingdom
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20110912
PL  - United States
TA  - PLoS One
JT  - PloS one
JID - 101285081
RN  - 0 (Homeobox Protein Nkx-2.5)
RN  - 0 (Homeodomain Proteins)
RN  - 0 (NKX2-5 protein, human)
RN  - 0 (PIAS1 protein, human)
RN  - 0 (PIAS2 protein, human)
RN  - 0 (Protein Inhibitors of Activated STAT)
RN  - 0 (SUMO-1 Protein)
RN  - 0 (SUMO2 protein, human)
RN  - 0 (Small Ubiquitin-Related Modifier Proteins)
RN  - 0 (Transcription Factors)
RN  - 0 (UBA2 protein, human)
RN  - EC 6.2.1.45 (Ubiquitin-Activating Enzymes)
SB  - IM
MH  - Animals
MH  - Blotting, Western
MH  - COS Cells
MH  - Cell Line
MH  - Chlorocebus aethiops
MH  - Electrophoretic Mobility Shift Assay
MH  - Fluorescent Antibody Technique
MH  - Homeobox Protein Nkx-2.5
MH  - Homeodomain Proteins/*metabolism
MH  - Humans
MH  - Immunohistochemistry
MH  - Immunoprecipitation
MH  - Mice
MH  - Myocardium/*metabolism
MH  - Protein Inhibitors of Activated STAT/metabolism
MH  - SUMO-1 Protein/genetics/*metabolism
MH  - Small Ubiquitin-Related Modifier Proteins/metabolism
MH  - Transcription Factors/*metabolism
MH  - Ubiquitin-Activating Enzymes/metabolism
PMC - PMC3171482
COIS- Competing Interests: The authors have declared that no competing interests exist.
EDAT- 2011/09/21 06:00
MHDA- 2012/03/01 06:00
PMCR- 2011/09/12
CRDT- 2011/09/21 06:00
PHST- 2011/03/14 00:00 [received]
PHST- 2011/08/22 00:00 [accepted]
PHST- 2011/09/21 06:00 [entrez]
PHST- 2011/09/21 06:00 [pubmed]
PHST- 2012/03/01 06:00 [medline]
PHST- 2011/09/12 00:00 [pmc-release]
AID - PONE-D-11-04674 [pii]
AID - 10.1371/journal.pone.0024812 [doi]
PST - ppublish
SO  - PLoS One. 2011;6(9):e24812. doi: 10.1371/journal.pone.0024812. Epub 2011 Sep 12.