Avian serum response factor expression restricted primarily to muscle cell lineages is required for alpha-actin gene transcription.
Serum response factor (SRF) gene expression in avian embryonic muscle lineages plays a central role in activating alpha-actin gene activity. In early stage HH 6 avian embryos, SRF mRNA expression showed strong localization to the neural groove, primitive streak, lateral plate mesoderm, and Hensen's node, while distinct SRF expression was seen later in the neural folds and the somites by HH stage 8. SRF transcripts appeared in the precardiac splanchnic mesoderm in stage HH 9 embryos and was detected at higher levels in the myocardium, somites, and lateral mesoderm of HH 11 embryos. SRF antibody staining demonstrated significant SRF protein accumulation in the myocardium of the developing heart and the myotomal portion of somites. During primary myogenesis in culture, SRF transcripts and nuclear SRF protein content increased about 40-fold, as primary myoblasts withdrew from the cell cycle, reaching their highest levels prior to the upregulation of the skeletal alpha-actin gene. A dominant-negative SRF mutant, SRFpm1, which inhibited DNA binding, but not dimerization of monomeric SRF subunits, blocked transcriptional activation of a skeletal alpha-actin promoter-luciferase reporter gene during myogenesis. Transcriptional blockade was reversed by co-transfections of a wild-type SRF expression vector, but was not rescued by the expression of other myogenic factors, such as MyoD and Mef-2C. Thus, SRF displayed an embryonic expression pattern restricted primarily to striated muscle cell lineages, in which increased mass of nuclear SRF was obligatory for alpha-actin gene transcription.
Croissant
J D
JD
Department of Cell Biology, Baylor College of Medicine, Houston, Texas 77030, USA.
Kim
J H
JH
Eichele
G
G
Goering
L
L
Lough
J
J
Prywes
R
R
Schwartz
R J
RJ
D000199:Actins / Q000235:genetics*; D000595:Amino Acid Sequence; D000818:Animals; D019070:Cell Lineage / Q000235:genetics; D002642:Chick Embryo; D018076:DNA, Complementary / Q000302:isolation & purification; D004268:DNA-Binding Proteins / Q000032:analysis / Q000235:genetics*; D018507:Gene Expression Regulation, Developmental; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D064326:MEF2 Transcription Factors; D008969:Molecular Sequence Data; D009132:Muscles / Q000737:chemistry / Q000196:embryology*; D017570:MyoD Protein / Q000235:genetics; D018006:Myogenic Regulatory Factors; D009687:Nuclear Proteins / Q000032:analysis / Q000235:genetics*; D011401:Promoter Regions, Genetic / Q000235:genetics; D012333:RNA, Messenger / Q000737:chemistry; D026362:Serum Response Factor; D014157:Transcription Factors / Q000032:analysis / Q000235:genetics*; D014158:Transcription, Genetic
D000199:Actins; D018076:DNA, Complementary; D004268:DNA-Binding Proteins; D064326:MEF2 Transcription Factors; D017570:MyoD Protein; D018006:Myogenic Regulatory Factors; D009687:Nuclear Proteins; D012333:RNA, Messenger; D026362:Serum Response Factor; D014157:Transcription Factors