Promoter upstream elements of the chicken cardiac myosin light-chain 2-A gene interact with trans-acting regulatory factors for muscle-specific transcription.

A segment of the 5'-flanking region of the chicken cardiac myosin light-chain gene extending from nucleotide -64 to the RNA start site is sufficient to allow muscle-specific transcription. In this paper, we characterize, by mutational analysis, sequence elements which are essential for the promoter activity. Furthermore, we present evidence for a negative-acting element which is possibly involved in conferring the muscle specificity. Nuclear proteins specifically bind to the DNA elements, as demonstrated by gel mobility shift assays and DNase I protection footprinting. The significance of the DNA-protein interactions for the function of the promoter in vivo is demonstrated by competition experiments in which protein-binding oligonucleotides were microinjected into nuclei of myotubes, where they successfully competed for the protein factors which are required to trans activate the MLC2-A promoter. The ability to bind nuclear proteins involves two closely spaced AT-rich sequence elements, one of which constitutes the TATA box. The binding properties correlate well with the capacity to activate transcription in vivo, since mutations in this region of the promoter concomitantly lead to loss of binding and transcriptional activity.

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Myosin light-chain 1 and 3 gene has two structurally distinct and differentially regulated promoters evolving at different rates

Molecular and Cellular Biology ◽

10.1128/mcb.5.11.3168-3182.1985 ◽

1985 ◽

Vol 5 (11) ◽

pp. 3168-3182

Author(s):

E E Strehler ◽

M Periasamy ◽

M A Strehler-Page ◽

B Nadal-Ginard

Keyword(s):

Light Chain ◽

Myosin Light Chain ◽

Mammalian Species ◽

In Vitro Transcription ◽

Chain Gene ◽

Promoter Regions ◽

Light Chain Gene ◽

Direct Initiation

DNA fragments located 10 kilobases apart in the genome and containing, respectively, the first myosin light chain 1 (MLC1f) and the first myosin light chain 3 (MLC3f) specific exon of the rat myosin light chain 1 and 3 gene, together with several hundred base pairs of upstream flanking sequences, have been shown in runoff in vitro transcription assays to direct initiation of transcription at the cap sites of MLC1f and MLC3f mRNAs used in vivo. These results establish the presence of two separate, functional promoters within that gene. A comparison of the nucleotide sequence of the rat MLC1f/3f gene with the corresponding sequences from mouse and chicken shows that: the MLC1f promoter regions have been highly conserved up to position -150 from the cap site while the MLC3f promoter regions display a very poor degree of homology and even the absence or poor conservation of typical eucaryotic promoter elements such as TATA and CAT boxes; the exon/intron structure of this gene has been completely conserved in the three species; and corresponding exons, except for the regions encoding most of the 5' and 3' untranslated sequences, show greater than 75% homology while corresponding introns are similar in size but considerably divergent in sequence. The above findings indicate that the overall structure of the MLC1f/3f genes has been maintained between avian and mammalian species and that these genes contain two functional and widely spaced promoters. The fact that the structures of the alkali light chain gene from Drosophila melanogaster and of other related genes of the troponin C supergene family resemble a MLC3f gene without an upstream promoter and first exon strongly suggests that the present-day MLC1f/3f genes of higher vertebrates arose from a primordial alkali light chain gene through the addition of a far-upstream MLC1f-specific promoter and first exon. The two promoters have evolved at different rates, with the MLC1f promoter being more conserved than the MLC3f promoter. This discrepant evolutionary rate might reflect different mechanisms of promoter activation for the transcription of MLC1f and MLC3f RNA.

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Myosin light-chain 1 and 3 gene has two structurally distinct and differentially regulated promoters evolving at different rates.

Molecular and Cellular Biology ◽

10.1128/mcb.5.11.3168 ◽

1985 ◽

Vol 5 (11) ◽

pp. 3168-3182 ◽

Cited By ~ 49

Author(s):

E E Strehler ◽

M Periasamy ◽

M A Strehler-Page ◽

B Nadal-Ginard

Keyword(s):

Light Chain ◽

Myosin Light Chain ◽

Mammalian Species ◽

In Vitro Transcription ◽

Chain Gene ◽

Promoter Regions ◽

Light Chain Gene ◽

Direct Initiation

DNA fragments located 10 kilobases apart in the genome and containing, respectively, the first myosin light chain 1 (MLC1f) and the first myosin light chain 3 (MLC3f) specific exon of the rat myosin light chain 1 and 3 gene, together with several hundred base pairs of upstream flanking sequences, have been shown in runoff in vitro transcription assays to direct initiation of transcription at the cap sites of MLC1f and MLC3f mRNAs used in vivo. These results establish the presence of two separate, functional promoters within that gene. A comparison of the nucleotide sequence of the rat MLC1f/3f gene with the corresponding sequences from mouse and chicken shows that: the MLC1f promoter regions have been highly conserved up to position -150 from the cap site while the MLC3f promoter regions display a very poor degree of homology and even the absence or poor conservation of typical eucaryotic promoter elements such as TATA and CAT boxes; the exon/intron structure of this gene has been completely conserved in the three species; and corresponding exons, except for the regions encoding most of the 5' and 3' untranslated sequences, show greater than 75% homology while corresponding introns are similar in size but considerably divergent in sequence. The above findings indicate that the overall structure of the MLC1f/3f genes has been maintained between avian and mammalian species and that these genes contain two functional and widely spaced promoters. The fact that the structures of the alkali light chain gene from Drosophila melanogaster and of other related genes of the troponin C supergene family resemble a MLC3f gene without an upstream promoter and first exon strongly suggests that the present-day MLC1f/3f genes of higher vertebrates arose from a primordial alkali light chain gene through the addition of a far-upstream MLC1f-specific promoter and first exon. The two promoters have evolved at different rates, with the MLC1f promoter being more conserved than the MLC3f promoter. This discrepant evolutionary rate might reflect different mechanisms of promoter activation for the transcription of MLC1f and MLC3f RNA.

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Tissue-specific DNase I-hypersensitive sites and hypomethylation in the chicken cardiac myosin light chain gene (L2-A).

Journal of Biological Chemistry ◽

10.1016/s0021-9258(19)76490-4 ◽

1987 ◽

Vol 262 (28) ◽

pp. 13750-13757

Author(s):

B B Winter ◽

H H Arnold

Keyword(s):

Light Chain ◽

Myosin Light Chain ◽

Dnase I ◽

Chain Gene ◽

Cardiac Myosin ◽

Light Chain Gene ◽

Tissue Specific ◽

Dnase I Hypersensitive Sites ◽

Hypersensitive Sites

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Cardiac Myosin Light Chain Kinase is Essential for Myosin Regulatory Light Chain Phosphorylation and Normal Cardiac Function in vivo

Biophysical Journal ◽

10.1016/j.bpj.2010.12.2203 ◽

2011 ◽

Vol 100 (3) ◽

pp. 369a

Author(s):

Peiguo Ding ◽

Jian Huang ◽

Pavan K. Battiprolu ◽

Joseph A. Hill ◽

Kristine E. Kamm ◽

...

Keyword(s):

Cardiac Function ◽

Light Chain ◽

Myosin Light Chain Kinase ◽

Myosin Light Chain ◽

Regulatory Light Chain ◽

Myosin Regulatory Light Chain ◽

Cardiac Myosin ◽

Regulatory Light Chain Phosphorylation ◽

Normal Cardiac Function

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Bom‐miR‐2805 upregulates the expression of Bombyx mori fibroin light chain gene in vivo

Journal of Cellular Biochemistry ◽

10.1002/jcb.28538 ◽

2019 ◽

Vol 120 (9) ◽

pp. 14326-14335

Author(s):

Yanhua Chen ◽

Tao Jiang ◽

Zhicheng Tan ◽

Peng Xue ◽

Jin Xu ◽

...

Keyword(s):

Bombyx Mori ◽

Light Chain ◽

Chain Gene ◽

Light Chain Gene ◽

Fibroin Light Chain

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Site-selected insertion of the transposon Tc1 into a Caenorhabditis elegans myosin light chain gene.

Molecular and Cellular Biology ◽

10.1128/mcb.13.2.902 ◽

1993 ◽

Vol 13 (2) ◽

pp. 902-910 ◽

Cited By ~ 42

Author(s):

A M Rushforth ◽

B Saari ◽

P Anderson

Keyword(s):

Polymerase Chain Reaction ◽

Caenorhabditis Elegans ◽

Light Chain ◽

Myosin Light Chain ◽

Null Alleles ◽

Chain Gene ◽

Splice Sites ◽

Chain Reaction ◽

Light Chain Gene ◽

Polymerase Chain

We used the polymerase chain reaction to detect insertions of the transposon Tc1 into mlc-2, one of two Caenorhabditis elegans regulatory myosin light chain genes. Our goals were to develop a general method to identify mutations in any sequenced gene and to establish the phenotype of mlc-2 loss-of-function mutants. The sensitivity of the polymerase chain reaction allowed us to identify nematode populations containing rare Tc1 insertions into mcl-2. mlc-2::Tc1 mutants were subsequently isolated from these populations by a sib selection procedure. We isolated three mutants with Tc1 insertions within the mlc-2 third exon and a fourth strain with Tc1 inserted in nearby noncoding DNA. To demonstrate the generality of our procedure, we isolated two additional mutants with Tc1 insertions within hlh-1, the C. elegans MyoD homolog. All of these mutants are essentially wild type when homozygous. Despite the fact that certain of these mutants have Tc1 inserted within exons of the target gene, these mutations may not be true null alleles. All three of the mlc-2 mutants contain mlc-2 mRNA in which all or part of Tc1 is spliced from the pre-mRNA, leaving small in-frame insertions or deletions in the mature message. There is a remarkable plasticity in the sites used to splice Tc1 from these mlc-2 pre-mRNAs; certain splice sites used in the mutants are very different from typical eukaryotic splice sites.

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