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rdf:type | |
lifeskim:mentions | |
pubmed:issue |
17
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pubmed:dateCreated |
1992-7-16
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pubmed:abstractText |
Segments of the 5'-flanking region of the mouse vascular smooth muscle alpha-actin gene were assayed for promoter activity in transfected mouse BC3H1 myogenic cells and AKR-2B embryonic fibroblasts. The region between -150 and -191 that functions as a positive transcriptional element in myogenic and fibroblastic cells contains a mammalian-specific inverted CC(A/T)6GG-type consensus sequence. Expression was restricted to fully differentiated myogenic cells when an additional sequence spanning -191 to -224 was included in reporter gene constructs. This 33-base pair (bp) negative regulatory element is 70% conserved between the mouse and human genes and contains a 10-bp motif at its 3' end that only partially resembles a CC(A/T)6GG element. Retention of a GGGA motif at the 3' boundary of the 33-bp region is sufficient to maintain full transcriptional repression in fibroblasts and is partly responsible for repression in undifferentiated myoblasts. Complete muscle tissue-restrictive expression requires an additional 8 bp from the CC(A/T)6GG-like element immediately 5' to the GGGA motif, since replacement of this region with an unrelated 10-bp sequence completely eliminated restrictive transcriptional behavior in undifferentiated myoblasts. The distal portion of the 5'-flanking region between -224 and -1074 contains six E-box motifs (CANNTG) and mediates high level transcription only in postconfluent BC3H1 myoblasts. Analysis of reporter gene constructs including either the proximal E-box at -240 or all six E-boxes indicate that the five distal E-boxes are not required for high level transcription. A 724-bp segment of the 5'-flanking region consisting of the proximal E-box flanked upstream by a mammalian-specific 352-bp region was sufficient for maximal transcriptional activation in postconfluent BC3H1 myoblasts. Deletion of the 352-bp region restricts the early transcriptional response to high cell density in temporal studies of promoter activity during BC3H1 myogenic cell differentiation.
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pubmed:grant | |
pubmed:language |
eng
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pubmed:journal | |
pubmed:citationSubset |
IM
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pubmed:chemical | |
pubmed:status |
MEDLINE
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pubmed:month |
Jun
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pubmed:issn |
0021-9258
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pubmed:author | |
pubmed:issnType |
Print
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pubmed:day |
15
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pubmed:volume |
267
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
11995-2003
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pubmed:dateRevised |
2008-11-21
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pubmed:meshHeading |
pubmed-meshheading:1601869-Actins,
pubmed-meshheading:1601869-Animals,
pubmed-meshheading:1601869-Base Sequence,
pubmed-meshheading:1601869-Cell Differentiation,
pubmed-meshheading:1601869-Chloramphenicol O-Acetyltransferase,
pubmed-meshheading:1601869-DNA,
pubmed-meshheading:1601869-Fibroblasts,
pubmed-meshheading:1601869-Gene Expression Regulation,
pubmed-meshheading:1601869-Growth Hormone,
pubmed-meshheading:1601869-Mice,
pubmed-meshheading:1601869-Molecular Sequence Data,
pubmed-meshheading:1601869-Muscle, Smooth, Vascular,
pubmed-meshheading:1601869-Plasmids,
pubmed-meshheading:1601869-Promoter Regions, Genetic,
pubmed-meshheading:1601869-Regulatory Sequences, Nucleic Acid,
pubmed-meshheading:1601869-Transcription, Genetic,
pubmed-meshheading:1601869-Transfection
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pubmed:year |
1992
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pubmed:articleTitle |
Positive and negative cis-acting regulatory elements mediate expression of the mouse vascular smooth muscle alpha-actin gene.
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pubmed:affiliation |
Ohio State Biochemistry Program, Ohio State University, Columbus 43210.
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pubmed:publicationType |
Journal Article,
Research Support, U.S. Gov't, P.H.S.,
Research Support, Non-U.S. Gov't
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