PMID- 18662983
OWN - NLM
STAT- MEDLINE
DCOM- 20081110
LR  - 20211020
IS  - 0021-9258 (Print)
IS  - 1083-351X (Electronic)
IS  - 0021-9258 (Linking)
VI  - 283
IP  - 39
DP  - 2008 Sep 26
TI  - Crossveinless-2 controls bone morphogenetic protein signaling during early 
      cardiomyocyte differentiation in P19 cells.
PG  - 26705-13
LID - 10.1074/jbc.M801485200 [doi]
AB  - Increasing evidence indicates that bone morphogenetic proteins (BMPs) are crucial 
      for cardiac induction, specification, and development. Although signaling of BMPs 
      is tightly regulated through soluble BMP-binding proteins, how they regulate BMP 
      signaling during cardiac differentiation remains unknown. To identify molecules 
      responsible for BMP signaling during early cardiomyocyte differentiation of P19 
      cells, cDNA subtraction was performed. We found a bimodal expression of the 
      BMP-binding protein Crossveinless-2 (Cv2) during cardiomyocyte differentiation; 
      Cv2 is temporally expressed earlier than cardiac transcription factors such as 
      Nkx2.5 and Tbx5 and acts as a suppressor for BMP signaling in P19 cells. We 
      established a P19 clonal cell line harboring a cardiac alpha-myosin heavy chain 
      promoter-driven enhanced green fluorescent protein gene to monitor cardiac 
      differentiation by flow cytometry. Treatment with BMP2 during the first 2 days of 
      differentiation suppressed cardiomyocyte differentiation through activation of 
      down-stream targets Smad1/5/8 protein and Id1 gene, whereas treatment with Cv2 
      conversely inhibited Smad1/5/8 activation and Id1 expression, leading to 
      increased generation of cardiac cells. RNA interference-mediated knockdown (KD) 
      of endogenous Cv2 showed increased Smad1/5/8 activation and impaired 
      cardiomyocyte differentiation. Expression of cardiac mesoderm markers was 
      reduced, whereas expression of Id1 and endoderm markers such as Sox7, Hnf4, and 
      E-cadherin was induced in Cv2-kinase dead cells. These phenotypes were rescued by 
      the addition of Cv2 protein to the culture media during the first 2 days of 
      differentiation or co-culture with parental cells. These data suggest that Cv2 
      may specify cardiac mesodermal lineage through inhibition of BMP signaling at 
      early stage of cardiogenesis.
FAU - Harada, Koichiro
AU  - Harada K
AD  - Department of Experimental Therapeutics, Translational Research Center, Kyoto 
      University Hospital, Kyoto 606-8507, Japan. kharada_kuhp@yahoo.co.jp
FAU - Ogai, Akiko
AU  - Ogai A
FAU - Takahashi, Tomosaburo
AU  - Takahashi T
FAU - Kitakaze, Masafumi
AU  - Kitakaze M
FAU - Matsubara, Hiroaki
AU  - Matsubara H
FAU - Oh, Hidemasa
AU  - Oh H
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20080728
PL  - United States
TA  - J Biol Chem
JT  - The Journal of biological chemistry
JID - 2985121R
RN  - 0 (Antigens, Differentiation)
RN  - 0 (Bmp2 protein, mouse)
RN  - 0 (Bone Morphogenetic Protein 2)
RN  - 0 (Bone Morphogenetic Proteins)
RN  - 0 (Carrier Proteins)
RN  - 0 (Muscle Proteins)
RN  - 0 (Transforming Growth Factor beta)
RN  - 0 (crossveinless 2 protein, mouse)
SB  - IM
MH  - Animals
MH  - Antigens, Differentiation/*biosynthesis/genetics
MH  - Bone Morphogenetic Protein 2
MH  - Bone Morphogenetic Proteins/*biosynthesis/genetics
MH  - Carrier Proteins/genetics/*metabolism
MH  - Cell Differentiation/*physiology
MH  - Cell Line, Tumor
MH  - Cell Lineage/physiology
MH  - Endoderm/cytology/embryology
MH  - Gene Expression Regulation, Developmental/*physiology
MH  - Mesoderm/cytology/embryology
MH  - Mice
MH  - Muscle Proteins/*biosynthesis/genetics
MH  - Myocytes, Cardiac/cytology/*metabolism
MH  - Organogenesis/physiology
MH  - Signal Transduction/*physiology
MH  - Transforming Growth Factor beta/*biosynthesis/genetics
PMC - PMC3258913
EDAT- 2008/07/30 09:00
MHDA- 2008/11/11 09:00
PMCR- 2009/09/26
CRDT- 2008/07/30 09:00
PHST- 2008/07/30 09:00 [pubmed]
PHST- 2008/11/11 09:00 [medline]
PHST- 2008/07/30 09:00 [entrez]
PHST- 2009/09/26 00:00 [pmc-release]
AID - S0021-9258(20)52413-7 [pii]
AID - 26705 [pii]
AID - 10.1074/jbc.M801485200 [doi]
PST - ppublish
SO  - J Biol Chem. 2008 Sep 26;283(39):26705-13. doi: 10.1074/jbc.M801485200. Epub 2008 
      Jul 28.