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Am J Physiol Renal Physiol 294: F525-F533, 2008. First published January 9, 2008; doi:10.1152/ajprenal.00299.2007
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Transcription of laminin {gamma}1 chain gene in rat mesangial cells: constitutive and inducible RNA polymerase II recruitment and chromatin states

Joel D. Nelson,1 Steve Flanagin,2 Yasunobu Kawata,2 Oleg Denisenko,2 and Karol Bomsztyk1,2

1Molecular and Cellular Biology Program and 2Department of Medicine, University of Washington Medicine Lake Union, Seattle, Washington

Submitted 29 June 2007 ; accepted in final form 2 January 2008

The laminin {gamma}1 chain, a critical component of the extracellular matrix, is encoded by the 125-kb-long Lamc1 locus. We profiled RNA polymerase II (Pol II) and histone modifications along the Lamc1 locus to explore transcription of this gene in its native chromatin environment. Treatment with 12-O-tetradecanoylphorbol-13-acetate increased Lamc1 mRNA in rat mesangial cells (RMC). This increase was matched by an increase in Pol II density along the entire length of the Lamc1 locus. In contrast, in the hepatocarcinoma cell line (HTC-IR) an increase in Pol II density was restricted to the promoter and was not followed by mRNA induction. The pattern of histone H3 methylation was similar for both cell types but an increase in H3 lysine 9 acetylation observed at the 5'-end was weaker in HTC-IR cells than in RMC. All of the histone modifications showed spatial patterns where levels differed greatly between the 5'- and 3'-ends of Lamc1. Conversely, at the short, highly induced egr-1 gene the differences in chromatin marks between the 5'- and 3'-ends were much smaller. The results of this study suggest that 1) Lamc1 transcription can be controlled after transcription initiation, to our knowledge, the first time this has been shown in an extracellular matrix gene, and 2) the length of a gene is a factor that can affect the chromatin environment for Pol II elongation.

Fast ChIP; extracellular matrix; hnRNP K; histone; elongation; egr-1


Address for reprint requests and other correspondence: K. Bomsztyk, UW Medicine at Lake Union, Box 358050, Univ. of Washington, Seattle, WA 98109 (e-mail: karolb{at}u.washington.edu)




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