The 5′-flanking region of the mouse vascular smooth muscle α-actin gene contains evolutionarily conserved sequence motifs within a functional promoter

Bon Hong Min, Douglas N. Foster, Arthur R. Strauch

Research output: Contribution to journalArticle

83 Citations (Scopus)

Abstract

The 5′-flanking, 5′-untranslated, and amino-terminal protein coding regions of the single-copy 13-kilobase mouse vascular smooth muscle (VSM) α-actin gene have been cloned and sequenced. Respectively, there is 73 and 89% homology from the start of transcription (+1) to a point 206 base pairs upstream when comparing mouse to chicken and mouse to human VSM α-actin 5′-flanking region sequences. Two proximal 16-base pair motifs containing putative cis- acting regulatory elements having the configuration CC(A/T)6GG were found to be 100% conserved and present in the same position upstream from the transcription start site in all three species. A third more distal CC(A/T)6GG-like motif was 100% conserved between only the mouse and human genes whereas a fourth motif was unique to the mouse gene. The two upstream motifs may be important in controlling VSM α-actin gene transcription in mammals. Cell transfection assays using hGH reporter gene fusion plasmids showed that all four CC(A/T)6GG elements were required for tissue-specific, core promoter activity and were able to direct hGH expression in both mouse BC3H1 myogenic cells and early-passage rabbit aortic smooth muscle cells. The core promoter was not active in mouse fibroblasts suggesting that the region between -372 and -143 may mediate tissue-restrictive expression of the VSM α-actin gene. A putative "cell density responsive element" may be located between -1074 and -372 since fusion plasmids containing this portion of the VSM α-actin 5′-flanking region were significantly more active in promoting hGH expression in inducible, density-activated BC3H1 myoblasts compared to aortic smooth muscle cells which are largely constitutive for VSM α-actin expression.

Original languageEnglish
Pages (from-to)16667-16675
Number of pages9
JournalJournal of Biological Chemistry
Volume265
Issue number27
Publication statusPublished - 1990 Sep 25
Externally publishedYes

Fingerprint

Conserved Sequence
5' Flanking Region
Vascular Smooth Muscle
Muscle
Actins
Genes
Base Pairing
Transcription
Smooth Muscle Myocytes
Plasmids
Fusion reactions
Cells
Tissue
Myoblasts
Transcription Initiation Site
Gene Fusion
Mammals
Reporter Genes
Fibroblasts
Open Reading Frames

ASJC Scopus subject areas

  • Biochemistry

Cite this

The 5′-flanking region of the mouse vascular smooth muscle α-actin gene contains evolutionarily conserved sequence motifs within a functional promoter. / Min, Bon Hong; Foster, Douglas N.; Strauch, Arthur R.

In: Journal of Biological Chemistry, Vol. 265, No. 27, 25.09.1990, p. 16667-16675.

Research output: Contribution to journalArticle

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