title: Leiomodin 1, a new serum response factor-dependent target gene expressed preferentially in differentiated smooth muscle cells.
abstract: Smooth muscle cell (SMC) differentiation is defined largely by a number of cell-restricted genes governed directly by the serum response factor (SRF)/myocardin (MYOCD) transcriptional switch. Here, we describe a new SRF/MYOCD-dependent, SMC-restricted gene known as Leiomodin 1 (Lmod1). Conventional and quantitative RT-PCRs indicate that Lmod1 mRNA expression is enriched in SMC-containing tissues of the mouse, whereas its two paralogs, Lmod2 and Lmod3, exhibit abundant expression in skeletal and cardiac muscle with very low levels in SMC-containing tissues. Western blotting and immunostaining of various adult and embryonic mouse tissues further confirm SMC-specific expression of the LMOD1 protein. Comparative genomic analysis of the human LMOD1 and LMOD2 genes with their respective mouse and rat orthologs shows high conservation between the three exons and several noncoding sequences, including the immediate 5' promoter region. Two conserved CArG boxes are present in both the LMOD1 and LMOD2 promoter regions, although LMOD1 displays much higher promoter activity and is more responsive to SRF/MYOCD stimulation. Gel shift assays demonstrate clear binding between SRF and the two CArG boxes in human LMOD1. Although the CArG boxes in LMOD1 and LMOD2 are similar, only LMOD1 displays SRF or MYOCD-dependent activation. Transgenic mouse studies reveal wild type LMOD1 promoter activity in cardiac and vascular SMC. Such activity is abolished upon mutation of both CArG boxes. Collectively, these data demonstrate that Lmod1 is a new SMC-restricted SRF/MYOCD target gene.
authors.affiliation: Aab Cardiovascular Research Institute, University of Rochester School of Medicine and Dentistry, Rochester, New York 14642, USA.
authors.forename: Vivek
authors.initials: V
authors.lastname: Nanda
authors.forename: Joseph M
authors.initials: JM
authors.lastname: Miano
mesh_terms: D000818:Animals; D001324:Autoantigens / Q000096:biosynthesis* / Q000235:genetics; D019556:COS Cells; D002454:Cell Differentiation / Q000502:physiology*; D002522:Chlorocebus aethiops; D003598:Cytoskeletal Proteins / Q000096:biosynthesis* / Q000235:genetics; D005786:Gene Expression Regulation / Q000502:physiology*; D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D051379:Mice; D018345:Mice, Knockout; D008840:Microfilament Proteins / Q000096:biosynthesis / Q000235:genetics; D009124:Muscle Proteins / Q000096:biosynthesis* / Q000235:genetics; D032389:Myocytes, Smooth Muscle / Q000166:cytology / Q000378:metabolism*; D041681:NIH 3T3 Cells; D009928:Organ Specificity / Q000502:physiology; D051381:Rats; D027602:Serum Response Element / Q000502:physiology*; D026362:Serum Response Factor / Q000235:genetics / Q000276:immunology*
chemical_list: D001324:Autoantigens; D003598:Cytoskeletal Proteins; C103926:LMOD1 protein, human; C000595329:LMOD3 protein, human; C545284:Lmod2 protein, rat; D008840:Microfilament Proteins; D009124:Muscle Proteins; C476858:SRF protein, human; D026362:Serum Response Factor; C523689:leiomodin protein, mouse