scholarly journals Bombyx mori miR-2845 represses the expression of fibroin light chain gene both in vitro and in vivo

PLoS ONE ◽  
2021 ◽  
Vol 16 (12) ◽  
pp. e0261391
Author(s):  
JingYi Huang ◽  
YanHua Chen ◽  
Juan Zhu ◽  
MeiXian Wang ◽  
ShunMing Tang ◽  
...  
Keyword(s):  
Bombyx Mori ◽  
Light Chain ◽  
Luciferase Reporter ◽  
Regulatory Function ◽  
Functional Verification ◽  
Chain Gene ◽  
Light Chain Gene ◽  
Expression Levels ◽  
Fibroin Light Chain

To study the regulatory function of Bombyx mori (B. mori) miRNAs (bmo-miR) on the expression of fibroin light chain gene (BmFib-L), the 3’UTR of BmFib-L mRNA was used as the target for online prediction of miRNAs from miRBase using RNAhybrid Software, and miR-2845 was screened out. First, the expression profiles of miR-2845 and BmFib-L in larvae of the 5th instar were analyzed by Real-time quantitative PCR (RT-qPCR). Then recombinant plasmids (pcDNA3.0-pre-miR-2845 and pGL3.0-BmFib-L) were constructed to use for the expression of miR-2845 and BmFib-L 3’UTR, respectively. Cellular-level functional verification of miR-2845 on BmFib-L was carried out using multiple experimental methods (including dual luciferase reporter vectors, artificially synthesized mimics and inhibitors, and target site mutations). Finally, in vivo functional verification was performed by injecting the recombinant vector in 5th instar larvae. BmFib-L expression levels were detected using RT-qPCR in the posterior silk glands (PSG) of the injected larvae. Results showed that the expression of miR-2845 increased between the 1st and 5th day in 5th instar larvae, but began to decline on the 5th day, while the expression of the target gene BmFib-L increased sharply. This suggests that miR-2845 and BmFib-L expression levels show opposing trends, implying a negative regulatory relationship. In BmN cells, miR-2845 significantly down-regulated the expression of BmFib-L; the inhibitory effect of miR-2845 on BmFib-L was disappeared after mutation of the targeting site on 3’UTR of BmFib-L; in individuals, miR-2845 significantly down-regulated BmFib-L expression levels. Our results provide new experimental data for clarifying the molecular regulation mechanism of silk protein expression.

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

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

scholarly journals Allelic variability in the third intron of the fibroin light chain gene in Bombyx mori (Lepidoptera: Bombycidae)

2009 ◽  
Vol 8 (1) ◽  
pp. 197-206 ◽  
Author(s):  
J.F. Barbosa ◽  
J.P. Bravo ◽  
D.B. Zanatta ◽  
J.L.C. Silva ◽  
M.A. Fernandez
Keyword(s):  
Bombyx Mori ◽  
Light Chain ◽  
Chain Gene ◽  
Light Chain Gene ◽  
The Third ◽  
Allelic Variability ◽  
Fibroin Light Chain

scholarly journals Linkage analysis of the fibroin light chain gene in the silkworm, Bombyx mori.

1984 ◽  
Vol 59 (3) ◽  
pp. 285-296 ◽  
Author(s):  
Akio HYODO ◽  
Toshio YAMAMOTO ◽  
Hitoshi UEDA ◽  
Fusaho TAKEI ◽  
Ken-ichi KIMURA ◽  
...  
Keyword(s):  
Linkage Analysis ◽  
Bombyx Mori ◽  
Light Chain ◽  
Chain Gene ◽  
Light Chain Gene ◽  
Fibroin Light Chain ◽  
Silkworm Bombyx Mori

scholarly journals bmo-miR-0001 and bmo-miR-0015 down-regulate expression of Bombyx mori fibroin light chain gene in vitro

2016 ◽  
Vol 17 (2) ◽  
pp. 127-135 ◽  
Author(s):  
Chen Chen ◽  
Yang-yang Fan ◽  
Xin Wang ◽  
Fei Song ◽  
Tao Jiang ◽  
...  
Keyword(s):  
Bombyx Mori ◽  
Light Chain ◽  
Chain Gene ◽  
Light Chain Gene ◽  
Fibroin Light Chain

scholarly journals Intrinsic bent DNA colocalizes with the sequence involved in the Nd-sD mutation in the Bombyx mori fibroin light chain gene

BMB Reports ◽  
2008 ◽  
Vol 41 (5) ◽  
pp. 394-399 ◽  
Author(s):  
Joice Felipes Barbosa ◽  
Juliana Pereira Bravo ◽  
Karen Izumi Takeda ◽  
Daniela Bertolini Zanatta ◽  
Jose Luis Da Conceicao Silva ◽  
...  
Keyword(s):  
Bombyx Mori ◽  
Light Chain ◽  
Chain Gene ◽  
Bent Dna ◽  
Light Chain Gene ◽  
Fibroin Light Chain

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

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.

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

1989 ◽  
Vol 9 (6) ◽  
pp. 2513-2525
Author(s):  
T Braun ◽  
E Tannich ◽  
G Buschhausen-Denker ◽  
H H Arnold
Keyword(s):  
Light Chain ◽  
Myosin Light Chain ◽  
Mutational Analysis ◽  
Nuclear Proteins ◽  
Chain Gene ◽  
Cardiac Myosin ◽  
Mobility Shift ◽  
Light Chain Gene ◽  
Sequence Elements

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.

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

1989 ◽  
Vol 9 (6) ◽  
pp. 2513-2525 ◽  
Author(s):  
T Braun ◽  
E Tannich ◽  
G Buschhausen-Denker ◽  
H H Arnold
Keyword(s):  
Light Chain ◽  
Myosin Light Chain ◽  
Mutational Analysis ◽  
Nuclear Proteins ◽  
Chain Gene ◽  
Cardiac Myosin ◽  
Mobility Shift ◽  
Light Chain Gene ◽  
Sequence Elements

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.

scholarly journals Phospholipase Cγ2 Contributes to Light-Chain Gene Activation and Receptor Editing

2007 ◽  
Vol 27 (17) ◽  
pp. 5957-5967 ◽  
Author(s):  
Li Bai ◽  
Yuhong Chen ◽  
Yinghong He ◽  
Xuezhi Dai ◽  
Xueyan Lin ◽  
...  
Keyword(s):  
B Cells ◽  
B Cell ◽  
Light Chain ◽  
Gene Activation ◽  
Chain Gene ◽  
Receptor Editing ◽  
Light Chain Gene ◽  
Transitional Type

ABSTRACT Phospholipase Cγ2 (PLCγ2) is critical for pre-B-cell receptor (pre-BCR) and BCR signaling. Current studies discovered that PLCγ2-deficient mice had reduced immunoglobulin λ (Igλ) light-chain usage throughout B-cell maturation stages, including transitional type 1 (T1), transitional type 2 (T2), and mature follicular B cells. The reduction of Igλ rearrangement by PLCγ2 deficiency was not due to specifically increased apoptosis or decreased proliferation of mutant Igλ+ B cells, as lack of PLCγ2 exerted a similar effect on apoptosis and proliferation of both Igλ+ and Igκ+ B cells. Moreover, PLCγ2-deficient IgHEL transgenic B cells exhibited an impairment of antigen-induced receptor editing among both the endogenous λ and κ loci in vitro and in vivo. Importantly, PLCγ2 deficiency impaired BCR-induced expression of IRF-4 and IRF-8, the two transcription factors critical for λ and κ light-chain rearrangements. Taken together, these data demonstrate that the PLCγ2 signaling pathway plays a role in activation of light-chain loci and contributes to receptor editing.

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