B-Myb Expression in Vascular Smooth Muscle Cells Occurs in a Cell Cycle-dependent Fashion and Down-regulates Promoter Activity of Type I Collagen Genes

Abstract

The members of the Myb family of transcription factors are defined by homology in the DNA-binding domain; all bind the Myb-binding site (MBS) sequence (YG(A/G)C(A/C/G)GTT(G/A)). Here we report that cultured bovine vascular smooth muscle cells (SMCs) express B-myb. Levels of B-myb RNA found in exponential growth were reduced dramatically in serum-deprived quiescent SMCs; B-myb mRNA levels increased in the cell cycle during the late G1 to S phase transition following restimulation with serum, epidermal growth factor, or phorbol ester plus insulin-like growth factor-1. Changes in the rate of B-myb gene transcription could account for part of the observed increase following serum addition. Treatment of SMC cultures with actinomycin D indicated a >4-h half-life for B-myb mRNA during the S phase of the cell cycle. Cotransfection of either a bovine or human B-myb expression vector down-regulated the activity of a multimerized MBS element-driven reporter construct in SMCs. Putative MBS elements were detected upstream of the promoters of the two chains of type I collagen, which we have found to be expressed inversely with growth state of the SMC (Kindy, M. S., Chang, C.-J., and Sonenshein, G. E. (1988) J. Biol. Chem. 263, 11426-11430). In cotransfection experiments, B-myb expression down-regulated the promoter activity of alpha1(I) and alpha2(I) collagen constructs an average of 92 and 82%, respectively. Thus, B-myb represents a potential link in the observed inverse relationship between collagen gene expression and growth of vascular SMCs.