PMID- 10082583
OWN - NLM
STAT- MEDLINE
DCOM- 19990420
LR  - 20190508
IS  - 0270-7306 (Print)
IS  - 0270-7306 (Linking)
VI  - 19
IP  - 4
DP  - 1999 Apr
TI  - cdk1- and cdk2-mediated phosphorylation of MyoD Ser200 in growing C2 myoblasts:
      role in modulating MyoD half-life and myogenic activity.
PG  - 3167-76
AB  - We have examined the role of protein phosphorylation in the modulation of the key
      muscle-specific transcription factor MyoD. We show that MyoD is highly
      phosphorylated in growing myoblasts and undergoes substantial dephosphorylation
      during differentiation. MyoD can be efficiently phosphorylated in vitro by either
      purified cdk1-cyclin B or cdk1 and cdk2 immunoprecipitated from proliferative
      myoblasts. Comparative two-dimensional tryptic phosphopeptide mapping combined
      with site-directed mutagenesis revealed that cdk1 and cdk2 phosphorylate MyoD on 
      serine 200 in proliferative myoblasts. In addition, when the seven
      proline-directed sites in MyoD were individually mutated, only substitution of
      serine 200 to a nonphosphorylatable alanine (MyoD-Ala200) abolished the
      slower-migrating hyperphosphorylated form of MyoD, seen either in vitro after
      phosphorylation by cdk1-cyclin B or in vivo following overexpression in 10T1/2
      cells. The MyoD-Ala200 mutant displayed activity threefold higher than that of
      wild-type MyoD in transactivation of an E-box-dependent reporter gene and
      promoted markedly enhanced myogenic conversion and fusion of 10T1/2 fibroblasts
      into muscle cells. In addition, the half-life of MyoD-Ala200 protein was longer
      than that of wild-type MyoD, substantiating a role of Ser200 phosphorylation in
      regulating MyoD turnover in proliferative myoblasts. Taken together, our data
      show that direct phosphorylation of MyoD Ser200 by cdk1 and cdk2 plays an
      integral role in compromising MyoD activity during myoblast proliferation.
FAU - Kitzmann, M
AU  - Kitzmann M
AD  - Institut de Genetique Humaine, Centre National de Recherche Scientifique, UPR
      1142, 34396 Montpellier cedex 5, France.
FAU - Vandromme, M
AU  - Vandromme M
FAU - Schaeffer, V
AU  - Schaeffer V
FAU - Carnac, G
AU  - Carnac G
FAU - Labbe, J C
AU  - Labbe JC
FAU - Lamb, N
AU  - Lamb N
FAU - Fernandez, A
AU  - Fernandez A
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Mol Cell Biol
JT  - Molecular and cellular biology
JID - 8109087
RN  - 0 (MyoD Protein)
RN  - 452VLY9402 (Serine)
RN  - EC 2.7.11.1 (Protein-Serine-Threonine Kinases)
RN  - EC 2.7.11.22 (CDC2 Protein Kinase)
RN  - EC 2.7.11.22 (CDC2-CDC28 Kinases)
RN  - EC 2.7.11.22 (Cyclin-Dependent Kinase 2)
RN  - EC 2.7.11.22 (Cyclin-Dependent Kinases)
RN  - OF5P57N2ZX (Alanine)
SB  - IM
MH  - Alanine/genetics/metabolism
MH  - CDC2 Protein Kinase/*metabolism
MH  - *CDC2-CDC28 Kinases
MH  - Cell Differentiation
MH  - Cell Division
MH  - Cyclin-Dependent Kinase 2
MH  - Cyclin-Dependent Kinases/*metabolism
MH  - Half-Life
MH  - Muscle, Skeletal/*cytology/metabolism
MH  - MyoD Protein/*metabolism
MH  - Phosphorylation
MH  - Protein-Serine-Threonine Kinases/*metabolism
MH  - Serine/metabolism
MH  - Stem Cells/*cytology/metabolism
MH  - Transcriptional Activation
PMC - PMC84110
EDAT- 1999/03/19 00:00
MHDA- 1999/03/19 00:01
CRDT- 1999/03/19 00:00
PHST- 1999/03/19 00:00 [pubmed]
PHST- 1999/03/19 00:01 [medline]
PHST- 1999/03/19 00:00 [entrez]
AID - 10.1128/mcb.19.4.3167 [doi]
PST - ppublish
SO  - Mol Cell Biol. 1999 Apr;19(4):3167-76. doi: 10.1128/mcb.19.4.3167.