Functional involvement of serum response factor in the transcriptional regulation of caldesmon gene.
A 22-bp fragment including the CArG element (CArG1) is essential for the transcription of the caldesmon gene. In this study, we investigated the effects of serum response factor (SRF) on the functional regulation of caldesmon promoter in smooth muscle cells. Gel supershift assay revealed that SRF was one component of the CArG1-protein complex. Dominant-negative mutants of SRF suppressed the promoter activity of caldesmon, whereas wild-type SRF overcame this suppression. These results suggest that SRF functions as a core activating factor of the caldesmon promoter. Furthermore, fractionation of smooth muscle cells' nuclear extracts using DNA affinity paramagnetic particles suggests that SRF transactivates the caldesmon promoter in concert with additional factors in the flow-through fraction recruited to the CArG element.
Momiyama
T
T
Department of Neurochemistry and Neuropharmacology, Biomedical Research Center, Osaka University Medical School, Japan.
Hayashi
K
K
Obata
H
H
Chimori
Y
Y
Nishida
T
T
Ito
T
T
Kamiike
W
W
Matsuda
H
H
Sobue
K
K
D000818:Animals; D015152:Blotting, Northern; D002148:Calmodulin-Binding Proteins / Q000235:genetics*; D002454:Cell Differentiation / Q000502:physiology; D002642:Chick Embryo; D004268:DNA-Binding Proteins / Q000032:analysis / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D005786:Gene Expression Regulation / Q000235:genetics*; D017930:Genes, Reporter / Q000235:genetics; D016297:Mutagenesis, Site-Directed / Q000235:genetics; D009687:Nuclear Proteins / Q000032:analysis / Q000302:isolation & purification / Q000378:metabolism*; D011401:Promoter Regions, Genetic / Q000235:genetics; D012333:RNA, Messenger / Q000032:analysis; D026362:Serum Response Factor; D015533:Transcriptional Activation / Q000235:genetics; D014162:Transfection / Q000235:genetics
D002148:Calmodulin-Binding Proteins; D004268:DNA-Binding Proteins; D009687:Nuclear Proteins; D012333:RNA, Messenger; D026362:Serum Response Factor