Disruption of the coordinate expression of muscle genes in a transfected BC3H1 myoblast cell line producing a low level of the adenovirus E1A transforming protein

1992 ◽  
Vol 70 (10-11) ◽  
pp. 1268-1276 ◽  
Author(s):  
Joe S. Mymryk ◽  
Janice D. Oakes ◽  
Senthil K. Muthuswamy ◽  
Pietro D'Amico ◽  
Stanley T. Bayley ◽  
...  
Keyword(s):  
Myogenic Differentiation ◽  
Calf Serum ◽  
Adenovirus E1a ◽  
Myoblast Cell Line ◽  
Muscle Genes ◽  
E1a Gene ◽  
H1 Cells ◽  
Myoblast Cell ◽  
Muscle Gene ◽  
Adenovirus 5

Mouse BC3H1 myoblasts were stably transfected with the adenovirus 5 E1A gene. One clonal line, BC3E7, was found to differ in some important respects from those previously reported for E1A-transformed myoblasts. In contrast to BC3H1 cells which differentiate when confluent in medium containing 0.5% fetal calf serum (FCS), BC3E7 cells failed to elongate and align, to express acetylcholine receptor and creatine kinase, and to down-regulate expression of β- and γ-actins and tropomyosin isoform (TM) 1. However, increased synthesis of TMs 2, 3, and 4, and myosin light chain 1 associated with differentiation in BC3H1 still occurred in BC3E7 cells, and most surprisingly, α-actin was produced at a significant level in both proliferating and confluent BC3E7 cells. Interestingly, myogenin was expressed in confluent BC3E7 cells in 0.5% FCS, but not in 20%. The level of E1A expression in BC3E7 cells was found to be very low by analysis of mRNA, by immunoprecipitation of E1A protein, and by the ability of BC3E7 cells to complement the E1A-deficient adenovirus mutant dl312. These results suggest that different levels of E1A may be needed to repress different promoters and that E1A does not block myogenic differentiation by repressing myogenin expression, but represses each muscle gene independently.Key words: actin, adenovirus 5 E1A, BC3H1 myoblasts, myogenin.

scholarly journals PFN2a Suppresses C2C12 Myogenic Development by Inhibiting Proliferation and Promoting Apoptosis via the p53 Pathway

Cells ◽  
2019 ◽  
Vol 8 (9) ◽  
pp. 959 ◽  
Author(s):  
Huaqin Li ◽  
Lianjie Hou ◽  
Yu Zhang ◽  
Fangyi Jiang ◽  
Yifan Zhu ◽  
...  
Keyword(s):  
Myogenic Differentiation ◽  
Inhibitory Effect ◽  
Cancer Growth ◽  
Actin Monomer ◽  
P53 Pathway ◽  
Histone Deacetylase 1 ◽  
Myoblast Cell Line ◽  
Myoblast Cell ◽  
Body Energy ◽  
Mouse Myoblast

Skeletal muscle plays a crucial role in physical activity and in regulating body energy and protein balance. Myoblast proliferation, differentiation, and apoptosis are indispensable processes for myoblast myogenesis. Profilin 2a (PFN2a) is a ubiquitous actin monomer-binding protein and promotes lung cancer growth and metastasis through suppressing the nuclear localization of histone deacetylase 1 (HDAC1). However, how PFN2a regulates myoblast myogenic development is still not clear. We constructed a C2C12 mouse myoblast cell line overexpressing PFN2a. The CRISPR/Cas9 system was used to study the function of PFN2a in C2C12 myogenic development. We find that PFN2a suppresses proliferation and promotes apoptosis and consequentially downregulates C2C12 myogenic development. The suppression of PFN2a also decreases the amount of HDAC1 in the nucleus and increases the protein level of p53 during C2C12 myogenic development. Therefore, we propose that PFN2a suppresses C2C12 myogenic development via the p53 pathway. Si-p53 (siRNA-p53) reverses the PFN2a inhibitory effect on C2C12 proliferation and the PFN2a promotion effect on C2C12 apoptosis, and then attenuates the suppression of PFN2a on myogenic differentiation. Our results expand understanding of PFN2a regulatory mechanisms in myogenic development and suggest potential therapeutic targets for muscle atrophy-related diseases.

A fourth human MEF2 transcription factor, hMEF2D, is an early marker of the myogenic lineage

Development ◽  
1993 ◽  
Vol 118 (4) ◽  
pp. 1095-1106 ◽  
Author(s):  
R.E. Breitbart ◽  
C.S. Liang ◽  
L.B. Smoot ◽  
D.A. Laheru ◽  
V. Mahdavi ◽  
...  
Keyword(s):  
Myogenic Differentiation ◽  
Specific Binding ◽  
Single Gene ◽  
Cell Types ◽  
Related Factor ◽  
Muscle Gene Expression ◽  
Molecular Events ◽  
Myogenic Lineage ◽  
Muscle Genes ◽  
Muscle Gene

The transition from multipotent mesodermal precursor to committed myoblast and its differentiation into a mature myocyte involve molecular events that enable the cell to activate muscle-specific genes. Among the participants in this process is the myocyte-specific enhancer factor 2 (MEF2) family of tissue-restricted transcription factors. These factors, which share a highly conserved DNA-binding domain including a MADS box, are essential for the expression of multiple muscle genes with cognate target MEF2 sites in cis. We report here a new human MEF2 factor, hMEF2D, which is unique among the members of this family in that it is present not only in myotubes but also in undifferentiated myoblasts, even before the appearance of myogenin. hMEF2D comprises several alternatively spliced products of a single gene, one of which is the human homolog of the Xenopus SRF-related factor SL-1. Like its relatives, cloned hMEF2D is capable of activating transcription via sequence-specific binding to the MEF2 site, recapitulating endogenous tissue-specific MEF2 activity. Indeed, while MEF2D mRNAs are ubiquitous, the protein is highly restricted to those cell types that contain this activity, implicating posttranscriptional mechanisms in the regulation of MEF2D expression. Alternative splicing may be important in this process: two alternative MEF2D domains, at least one of which is specifically included during myogenic differentiation, also correlate precisely with endogenous MEF2 activity. These findings provide compelling evidence that MEF2D is an integral link in the regulatory network for muscle gene expression. Its presence in undifferentiated myoblasts further suggests that it may be a mediator of commitment in the myogenic lineage.

scholarly journals Nanocomposites Based on PLLA and Multi Walled Carbon Nanotubes Support the Myogenic Differentiation of Murine Myoblast Cell Line

2013 ◽  
Vol 2013 ◽  
pp. 1-8 ◽  
Author(s):  
Simona Montesano ◽  
Erlantz Lizundia ◽  
Francesco D'Angelo ◽  
Elena Fortunati ◽  
Samantha Mattioli ◽  
...  
Keyword(s):  
Carbon Nanotubes ◽  
Myogenic Differentiation ◽  
Nanocomposite Materials ◽  
Neat Plla ◽  
Myoblast Cell Line ◽  
Multi Walled Carbon Nanotubes ◽  
Myoblast Cell ◽  
Walled Carbon Nanotubes ◽  
Myogenic Markers ◽  
Fusion Index

We explored the effect of poly(L-lactic acid) (PLLA) containing various percentages (0.1, 0.5, 1, and 3 wt.%) of multi walled carbon nanotubes (MWCNTs) on the myogenic differentiation of C2C12 murine myoblast progenitor cells. We showed that all PLLA/MWCNTs nanocomposite materials support the myotubes formation more efficiently than neat PLLA as indicated by the high expression of the most significant myogenic markers: MyoD, Myosin Heavy Chain, dimension of myofibres, and fusion myogenic index. Interestingly, we note that both MyoD and myogenic fusion index levels were in the order 0.1 MWCNTs = 0.5 MWCNTs > 1 MWCNTs > 3 MWCNTs > neat PLLA, suggesting that the amount of MWCNTs influenced the cell differentiation.

scholarly journals FLNC Expression Level Influences the Activity of TEAD-YAP/TAZ Signaling

Genes ◽  
2020 ◽  
Vol 11 (11) ◽  
pp. 1343
Author(s):  
Anastasia Knyazeva ◽  
Aleksandr Khudiakov ◽  
Raquel Vaz ◽  
Aleksey Muravyev ◽  
Ksenia Sukhareva ◽  
...  
Keyword(s):  
Muscle Cell ◽  
Myogenic Differentiation ◽  
Clinical Importance ◽  
Actin Binding ◽  
Hippo Signaling ◽  
Cell Functions ◽  
Myoblast Cell Line ◽  
Myoblast Cell ◽  
Migration Capacity

Filamin C (FLNC), being one of the major actin-binding proteins, is involved in the maintenance of key muscle cell functions. Inherited skeletal muscle and cardiac disorders linked to genetic variants in FLNC have attracted attention because of their high clinical importance and possibility of genotype-phenotype correlations. To further expand on the role of FLNC in muscle cells, we focused on detailed alterations of muscle cell properties developed after the loss of FLNC. Using the CRISPR/Cas9 method we generated a C2C12 murine myoblast cell line with stably suppressed Flnc expression. FLNC-deficient myoblasts have a significantly higher proliferation rate combined with an impaired cell migration capacity. The suppression of Flnc expression leads to inability to complete myogenic differentiation, diminished expression of Myh1 and Myh4, alteration of transcriptional dynamics of myogenic factors, such as Mymk and Myog, and deregulation of Hippo signaling pathway. Specifically, we identified elevated basal levels of Hippo activity in myoblasts with loss of FLNC, and ineffective reduction of Hippo signaling activity during myogenic differentiation. The latter was restored by Flnc overexpression. In summary, we confirmed the role of FLNC in muscle cell proliferation, migration and differentiation, and demonstrated for the first time the direct link between Flnc expression and activity of TEAD-YAP\TAZ signaling. These findings support a role of FLNC in regulation of essential muscle processes relying on mechanical as well as signaling mechanisms.

scholarly journals HIRA stabilizes skeletal muscle lineage identity

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Joana Esteves de Lima ◽  
Reem Bou Akar ◽  
Léo Machado ◽  
Yuefeng Li ◽  
Bernadette Drayton-Libotte ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Cell Lineage ◽  
Promoter Regions ◽  
Cell Fates ◽  
Regulatory Regions ◽  
Muscle Stem Cells ◽  
Adult Mice ◽  
H3k4me3 Mark ◽  
Muscle Genes ◽  
Muscle Gene

AbstractThe epigenetic mechanisms coordinating the maintenance of adult cellular lineages and the inhibition of alternative cell fates remain poorly understood. Here we show that targeted ablation of the histone chaperone HIRA in myogenic cells leads to extensive transcriptional modifications, consistent with a role in maintaining skeletal muscle cellular identity. We demonstrate that conditional ablation of HIRA in muscle stem cells of adult mice compromises their capacity to regenerate and self-renew, leading to tissue repair failure. Chromatin analysis of Hira-deficient cells show a significant reduction of histone variant H3.3 deposition and H3K27ac modification at regulatory regions of muscle genes. Additionally, we find that genes from alternative lineages are ectopically expressed in Hira-mutant cells via MLL1/MLL2-mediated increase of H3K4me3 mark at silent promoter regions. Therefore, we conclude that HIRA sustains the chromatin landscape governing muscle cell lineage identity via incorporation of H3.3 at muscle gene regulatory regions, while preventing the expression of alternative lineage genes.

scholarly journals Concentration dependence of transcriptional transactivation in inducible E1A-containing human cells.

1988 ◽  
Vol 8 (11) ◽  
pp. 4799-4807 ◽  
Author(s):  
L J Brunet ◽  
A J Berk
Keyword(s):  
Adenovirus Type ◽  
In Vitro Transcription ◽  
Mrna Levels ◽  
Adenovirus E1a ◽  
Transcription System ◽  
Nuclear Extracts ◽  
Tumor Virus ◽  
E1a Gene ◽  
High Level

The adenovirus E1A proteins are essential for the normal temporal activation of transcription from every other adenoviral early promoter. High-level E1A expression in the absence of viral infection would facilitate biochemical studies of E1A-mediated transactivation. Toward this end, we introduced the adenovirus type 2 E1A gene under the control of the murine mammary tumor virus promoter into HeLa cells. Uninduced cells expressed little or no detectable E1A mRNA. Upon induction, mRNA levels accumulated to about 50% of the level observed in 293 cells. The level of E1A expression in these cells could be controlled by varying the concentration of the inducing glucocorticoid. Under these conditions of varying E1A concentrations, it was observed that activation of the E2, E3, and E4 promoters of H5dl312 initiated at the same E1A concentration and that transcription from each promoter increased as the E1A concentration increased. These results indicate that E1A-mediated transactivation is proportional to the concentration of E1A protein. E1A-dependent transcriptional stimulation of the E4 promoter was reproduced in an in vitro transcription system, demonstrating that expression of only the E1A proteins was sufficient to increase the transcriptional activity of nuclear extracts.

scholarly journals Lentivector-Mediated Transfer of Bmi-1 and Telomerase in Muscle Satellite Cells Yields a Duchenne Myoblast Cell Line with Long-Term Genotypic and Phenotypic Stability

2003 ◽  
Vol 14 (16) ◽  
pp. 1525-1533 ◽  
Author(s):  
Christophe Cudré-Mauroux ◽  
Teresa Occhiodoro ◽  
Stéphane König ◽  
Patrick Salmon ◽  
Laurent Bernheim ◽  
...  
Keyword(s):  
Cell Line ◽  
Satellite Cells ◽  
Phenotypic Stability ◽  
Muscle Satellite Cells ◽  
Myoblast Cell Line ◽  
Myoblast Cell

Concentration dependence of transcriptional transactivation in inducible E1A-containing human cells

1988 ◽  
Vol 8 (11) ◽  
pp. 4799-4807
Author(s):  
L J Brunet ◽  
A J Berk
Keyword(s):  
Adenovirus Type ◽  
In Vitro Transcription ◽  
Mrna Levels ◽  
Adenovirus E1a ◽  
Transcription System ◽  
Nuclear Extracts ◽  
Tumor Virus ◽  
E1a Gene ◽  
High Level

The adenovirus E1A proteins are essential for the normal temporal activation of transcription from every other adenoviral early promoter. High-level E1A expression in the absence of viral infection would facilitate biochemical studies of E1A-mediated transactivation. Toward this end, we introduced the adenovirus type 2 E1A gene under the control of the murine mammary tumor virus promoter into HeLa cells. Uninduced cells expressed little or no detectable E1A mRNA. Upon induction, mRNA levels accumulated to about 50% of the level observed in 293 cells. The level of E1A expression in these cells could be controlled by varying the concentration of the inducing glucocorticoid. Under these conditions of varying E1A concentrations, it was observed that activation of the E2, E3, and E4 promoters of H5dl312 initiated at the same E1A concentration and that transcription from each promoter increased as the E1A concentration increased. These results indicate that E1A-mediated transactivation is proportional to the concentration of E1A protein. E1A-dependent transcriptional stimulation of the E4 promoter was reproduced in an in vitro transcription system, demonstrating that expression of only the E1A proteins was sufficient to increase the transcriptional activity of nuclear extracts.

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