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1 Pulmonary and Critical Care Division and Department of Medicine, Brigham and Women's Hospital, Boston, MA, USA; School of Biology, Kaohsiung Medical University, Kaohsiung, Taiwan, Taiwan
2 Pulmonary and Critical Care Division and Department of Medicine, Brigham and Women's Hospital, Boston, MA, USA
3 Pulmonary and Critical Care Division and Department of Medicine, Brigham and Women's Hospital, Boston, MA, USA; Division of Infectious Diseases and Department of Internal Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan, Taiwan
* To whom correspondence should be addressed. E-mail: syet{at}rics.bwh.harvard.edu.
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 transcription of CSRP2 in VSMC, we analyzed the 5'-flanking sequence of the CSRP2 gene. We showed previously that 4855 bp of 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 were generated and analyzed. Similar to that observed in the -4855CSRP2-lacZ mice, 
-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 -573 to -550 bp 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 (DN-SRF) did not repress CSRP2 promoter activity, which was different from the repressive effect that DN-SRF has on the SM22
promoter. Our data suggest that VSMC-specific element(s) are present within -573 to -550 bp of the CSRP2 5'-flanking sequence; however, in contrast to many other smooth muscle genes, the transcriptional regulation of CSRP2 gene is not dependent on SRF.
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