Identification of a CArG-independent region of the cysteine-rich protein 2 promoter that directs expression in the developing vasculature.
Cysteine-rich protein (CRP)2 is a member of the LIM-only CRP family that is expressed in vascular smooth muscle cells (VSMC). To gain insight into the transcription of CSRP2 (gene name for CRP2) in VSMC, we analyzed the 5'-flanking sequence of the CSRP2 gene. We showed previously that 4,855 bp of the 5'-flanking sequence of the CSRP2 gene directed lacZ reporter gene expression, primarily in the VSMC of transgenic mice. To further define the regulatory sequences important for CSRP2 expression in VSMC, a series of promoter constructs containing deletions of the 5'-flanking sequence upstream of a nuclear-localized lacZ reporter gene were generated and analyzed. Similar to that observed in the -4855CSRP2-lacZ mice, beta-galactosidase reporter activity was detected in the developing great vessels, aorta, intersegmental arteries, umbilical vessels, endocardial cushions, and neural tube in the -3513-, -2663-, -795-, and -664CSRP2-lacZ lines. However, an internal deletion of bp -573 to -550 abolished the vascular, but not the neural tube, staining. Interestingly, no CArG box [CC(A/T)6GG] was present in the -795-bp fragment. Cotransfection experiments showed that dominant-negative serum response factor (SRF) did not repress CSRP2 promoter activity, which was different from the repressive effect of dominant-negative SRF on the SM22 alpha promoter. Our data suggest the presence of a VSMC-specific element(s) within bp -573 to -550 of the CSRP2 5'-flanking sequence; however, in contrast to many other smooth muscle genes, transcriptional regulation of the CSRP2 gene is not dependent on SRF.
Chang
Yung-Fu
YF
Pulmonary and Critical Care Division, Department of Medicine, Brigham and Women's Hospital and Harvard Medical School, 75 Francis St., Boston, MA 02115, USA.
Wei
Jiao
J
Liu
Xiaoli
X
Chen
Yen-Hsu
YH
Layne
Matthew D
MD
Yet
Shaw-Fang
SF
D000375:Aging / Q000378:metabolism; D000818:Animals; D001483:Base Sequence / Q000235:genetics; D001808:Blood Vessels / Q000196:embryology* / Q000254:growth & development* / Q000378:metabolism; D018797:Cell Cycle Proteins; D002478:Cells, Cultured; D004268:DNA-Binding Proteins / Q000502:physiology; D004622:Embryo, Mammalian / Q000378:metabolism; D005314:Embryonic and Fetal Development; D015870:Gene Expression; D034481:Heterogeneous-Nuclear Ribonucleoprotein Group A-B; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000378:metabolism; D032389:Myocytes, Smooth Muscle / Q000378:metabolism; D009687:Nuclear Proteins / Q000235:genetics* / Q000378:metabolism; D011401:Promoter Regions, Genetic / Q000235:genetics / Q000502:physiology*; D017434:Protein Structure, Tertiary / Q000235:genetics; D011506:Proteins; D051381:Rats; D017207:Rats, Sprague-Dawley; D012097:Repressor Proteins / Q000502:physiology; D012261:Ribonucleoproteins; D014157:Transcription Factors; D019076:Transgenes
D018797:Cell Cycle Proteins; D004268:DNA-Binding Proteins; D034481:Heterogeneous-Nuclear Ribonucleoprotein Group A-B; C121363:Hnrnpab protein, rat; D009687:Nuclear Proteins; D011506:Proteins; D012097:Repressor Proteins; D012261:Ribonucleoproteins; D014157:Transcription Factors; C074370:cysteine-rich protein, mammalian; C077048:Hnrnpab protein, mouse