The nucleotide sequence of a rat myosin light chain 2 gene

Nudel, Uri; Calvo, Joseph M.; Shani, Moshe; Levy, Zehava

doi:10.1093/nar/12.18.7175

Cited by 82 publications

(35 citation statements)

References 42 publications

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“…It is interesting to note that an AUUUA repeated motif has been identified in several mitogen-regulated transcripts, in which it has been postulated to influence mRNA turnover (10). Also, a consensus sequence for recognition by the transcription factor AP-1 (25) (44), SV40 TC-II and SphI motifs (11), CArG motif (38), AP-2 binding site (39), IgH enhancer (2), and IgK enhancer (51). VOL.…”

Section: Resultsmentioning

confidence: 99%

Identification of upstream and intragenic regulatory elements that confer cell-type-restricted and differentiation-specific expression on the muscle creatine kinase gene.

Sternberg¹,

Spizz²,

Perry³

et al. 1988

Mol. Cell. Biol.

250

207

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Terminal differentiation of skeletal myoblasts is accompanied by induction of a series of tissue-specific gene products, which includes the muscle isoenzyme of creatine kinase (MCK). To begin to define the sequences and signals involved in MCK regulation in developing muscle cells, the mouse MCK gene has been isolated. Sequence analysis of 4,147 bases of DNA surrounding the transcription initiation site revealed several interesting structural features, some of which are common to other muscle-specific genes and to cellular and viral enhancers. To test for sequences required for regulated expression, a region upstream of the MCK gene from -4800 to +1 base pairs, relative to the transcription initiation site, was linked to the coding sequences of the bacterial chloramphenicol acetyltransferase (CAT) gene. Introduction of this MCK-CAT fusion gene into C2 muscle cells resulted in high-level expression of CAT activity in differentiated myotubes and no detectable expression in proliferating undifferentiated myoblasts or in nonmyogenic cell lines. Deletion mutagenesis of sequences between -4800 and the transcription start site showed that the region between -1351 and -1050 was sufficient to confer cell type-specific and developmentally regulated expression on the MCK promoter. This upstream regulatory element functioned independently of position, orientation, or distance from the promoter and therefore exhibited the properties of a classical enhancer. This upstream enhancer also was able to confer muscle-specific regulation on the simian virus 40 promoter, although it exhibited a 3- to 5-fold preference for its own promoter. In contrast to the cell type- and differentiation-specific expression of the upstream enhancer, the MCK promoter was able to function in myoblasts and myotubes and in nonmyogenic cell lines when combined with the simian virus 40 enhancer. An additional positive regulatory element was identified within the first intron of the MCK gene. Like the upstream enhancer, this intragenic element functioned independently of position, orientation, and distance with respect to the MCK promoter and was active in differentiated myotubes but not in myoblasts. These results demonstrate that expression of the MCK gene in developing muscle cells is controlled by complex interactions among multiple upstream and intragenic regulatory elements that are functional only in the appropriate cellular context.

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Section: Resultsmentioning

confidence: 99%

Identification of upstream and intragenic regulatory elements that confer cell-type-restricted and differentiation-specific expression on the muscle creatine kinase gene.

Sternberg¹,

Spizz²,

Perry³

et al. 1988

Mol. Cell. Biol.

250

207

View full text Add to dashboard Cite

show abstract

“…4B). iith BamHu was probed with nick-rat LC1, 805 nt (47); rat LC3, 167 nt (47); rat LC2, 236 nt analysis; the two bands are those (36); mouse LC1, 115 nt (42); and mouse LC3, about 120 nt (Fig. 1).…”

Section: Resultsmentioning

confidence: 99%

“…Although these elements may not prove to be sufficient to allow expression of heterologous structural genes in muscle, their functional significance is suggested by their repeated appearance in close proximity to muscle-specific transcription initiation sites and by their conservation between chicken and rat DNAs. These elements do not, however, seem to be necessary for the expression of all musclespecific genes, since they are not found within the available upstream sequences of the myosin light-chain genes (36,42,47).…”

Section: Discussionmentioning

confidence: 99%

Transcriptional regulation of the muscle creatine kinase gene and regulated expression in transfected mouse myoblasts.

Jaynes¹,

Chamberlain²,

Buskin³

et al. 1986

Mol. Cell. Biol.

179

122

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The muscle-specific form of creatine kinase (MCK) is induced in differentiating myoblast cultures, and a dramatic increase in mRNA levels precedes and parallels the increase in MCK protein. To study this induction, the complete MCK gene was cloned and characterized. The transcription unit was shown to span 11 kilobases and to contain seven introns. The splice junctions were identified and shown to conform to the appropriate consensus sequences. Close homology with branchpoint consensuses was found upstream of the 3' splice sites in six of seven cases. Transcriptional regulation of the gene in differentiating myoblast cultures was demonstrated by nuclear run-on experiments; increases in transcription accounted for a major part of the increased mRNA levels. Regulated expression of a transfected MCK gene containing the entire transcription unit with 3.3 kilobases of 5'-flanking sequence was also demonstrated during differentiation of the MM14 mouse myoblast cell line. The MCK 5'-flanking region was sufficient to confer transcriptional regulation to a heterologous structural gene, since chloramphenicol acetyl transferase activity was induced during differentiation of cultures transfected with an MCK-chloramphenicol acetyl transferase fusion construct. Examination of the DNA sequence immediately upstream of the transcription start site revealed a 17-nucleotide element which occurred three times. Comparisons with other muscle-specific genes which are also transcriptionally regulated during myogenesis revealed upstream homologies in the ca-actin and myosin heavy chain genes, but not in the myosin light-chain genes, with the regions containing these repeats. We suggest that coordinate control of a subset of muscle genes may occur via recognition of these common sequences.

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“…For example, the ANF promoter contains, in addition to the NKE, WGATAR, CArG, and AT-rich/Mef2 elements that may function in different combinatorial pathways to direct proper spatial and stage-specific transcription of the ANF gene (discussed below). A search for different cardiac promoters revealed the presence of NKE-like elements within the upstream regulatory regions of the cardiac myosin light-chain 2 (MLC2) (40) and ␤-myosin heavy-chain (␤MHC) genes (28) ( Table 2). Incidentally, expression of these two genes is altered in mice homozygous for a null Nkx-2.5 allele, which display defective chamber specification (31).…”

Section: Discussionmentioning

confidence: 99%

The Atrial Natriuretic Factor Promoter Is a Downstream Target for Nkx-2.5 in the Myocardium

Durocher

Chen

Ardati

et al. 1996

Molecular and Cellular Biology

144

106

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The recently described NK2 family of homeodomain proteins are key developmental regulators. In Drosophila melanogaster, two members of this family, bagpipe and tinman, are required for visceral and cardiac mesoderm formation, respectively. In vertebrates, tinman appears to represent a family of closely related NK2 genes, including Nkx-2.5, that are expressed at an early stage in precardiac cells. Consistent with a role for Nkx-2.5 in heart development, inactivation of the Nkx-2.5 gene in mice causes severe cardiac malformations and embryonic lethality. However, little is known about the molecular action of Nkx-2.5 and its targets in cardiac muscle. In this paper, we report the identification and characterization of a functional and highly conserved Nkx-2.5 response element, termed the NKE, in the proximal region of the cardiac atrial natriuretic factor (ANF) promoter. The NKE is composed of two near-consensus NK2 binding sites that are each able to bind purified Nkx-2.5. The NKE is sufficient to confer cardiac cell-specific activity to a minimal TATAcontaining promoter and is required for Nkx-2.5 activation of the ANF promoter in heterologous cells. Interestingly, in primary cardiocyte cultures, the NKE contributes to ANF promoter activity in a chamber-and developmental stage-specific manner, suggesting that Nkx-2.5 and/or other related cardiac proteins may play a role in chamber specification. This work provides the identification of a direct target for NK2 homeoproteins in the heart and lays the foundation for further molecular analyses of the role of Nkx-2.5 and other NK2 proteins in cardiac development.

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The nucleotide sequence of a rat myosin light chain 2 gene

Cited by 82 publications

References 42 publications

Identification of upstream and intragenic regulatory elements that confer cell-type-restricted and differentiation-specific expression on the muscle creatine kinase gene.

Identification of upstream and intragenic regulatory elements that confer cell-type-restricted and differentiation-specific expression on the muscle creatine kinase gene.

Transcriptional regulation of the muscle creatine kinase gene and regulated expression in transfected mouse myoblasts.

The Atrial Natriuretic Factor Promoter Is a Downstream Target for Nkx-2.5 in the Myocardium

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