Role of myofibril-inducing RNA in cardiac TnT expression in developing Mexican axolotl

Sferrazza, Gian Franco; Zhang, Chi; Jia, Pingping; Lemanski, Sharon L.; Athauda, Gagani; Stassi, Alyssa; Halager, Kristine; Maier, Jennifer; Rueda-de-Leon, Elena; Gupta, Amit; Dube, Syamalima; Huang, Xupei; Prentice, Howard; Dube, Dipak K.; Lemanski, Larry F.

doi:10.1016/j.bbrc.2007.03.064

Cited by 6 publications

(4 citation statements)

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“…A mechanism whereby the MIR enters the mutant heart cells, is transported into the nucleus, perhaps together with its binding protein(s) [ 7 ], and promotes gene transcription of the various myofibril components. The MIR could as well regulate RNA splicing processes for these genes as indicated in our studies on cardiac troponin T [ 23 ]. The myofibrillogenesis is thus promoted with increased expression of building blocks of myofibrillar proteins.…”

Section: Discussionmentioning

confidence: 99%

“…Reverse transcription is performed using the Thermoscript RT system from Invitrogen, CA. Quantitative PCR is performed in a Lightcycler system using a Roche's Fast Start SYBR Green I Kit, following our published methods [ 23 ]. The primers were designed by the Primer 3 program from the Massachusetts Institute of Technology as listed in the Table 1 .…”

Section: Methodsmentioning

confidence: 99%

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Myofibril-Inducing RNA (MIR) is essential for tropomyosin expression and myofibrillogenesis in axolotl hearts

et al. 2009

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The Mexican axolotl, Ambystoma mexicanum, carries the naturally-occurring recessive mutant gene 'c' that results in a failure of homozygous (c/c) embryos to form hearts that beat because of an absence of organized myofibrils. Our previous studies have shown that a noncoding RNA, Myofibril-Inducing RNA (MIR), is capable of promoting myofibrillogenesis and heart beating in the mutant (c/c) axolotls. The present study demonstrates that the MIR gene is essential for tropomyosin (TM) expression in axolotl hearts during development. Gene expression studies show that mRNA expression of various tropomyosin isoforms in untreated mutant hearts and in normal hearts knocked down with double-stranded MIR (dsMIR) are similar to untreated normal. However, at the protein level, selected tropomyosin isoforms are significantly reduced in mutant and dsMIR treated normal hearts. These results suggest that MIR is involved in controlling the translation or post-translation of various TM isoforms and subsequently of regulating cardiac contractility.

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Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Myofibril-Inducing RNA (MIR) is essential for tropomyosin expression and myofibrillogenesis in axolotl hearts

et al. 2009

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“…Real time RT-PCR experiments were performed according to our published procedures [ 16 , 17 ]. The primer sequences used in these studies were as follows:…”

Section: Methodsmentioning

confidence: 99%

Identification of a truncated form of methionine sulfoxide reductase a expressed in mouse embryonic stem cells

et al. 2011

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BackgroundMethionine Sulfoxide Reductase A (MsrA), an enzyme in the Msr gene family, is important in the cellular anti-oxidative stress defense mechanism. It acts by reducing the oxidized methionine sulfoxide in proteins back to sulfide and by reducing the cellular level of reactive oxygen species. MsrA, the only enzyme in the Msr gene family that can reduce the S-form epimers of methionine sulfoxide, has been located in different cellular compartments including mitochondria, cytosol and nuclei of various cell lines.MethodsIn the present study, we have isolated a truncated form of the MsrA transcript from cultured mouse embryonic stem cells and performed eGFP fusion protein expression, confocal microscopy and real time RT-PCR studies.ResultsResults show a different expression response of this truncated transcript to oxygen deprivation and reoxygenation treatments in stem cells, compared to the longer full length form. In addition, a different subcellular localization pattern was noted with most of the eGFP fusion protein detected in the cytosol.ConclusionOne possibility for the existence of a truncated form of the MsrA transcripts could be that with a smaller protein size, yet retaining a GCWFG action site, this protein might have easier access to oxidize methionine residues on proteins than the longer form of the MsrA protein, thus having an evolutionary selection advantage. This research opens the door for further study on the role and function of the truncated MsrA embryonic mouse stem cells.

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“…Animals carrying homozygous recessive alleles for c lack organized cardiac myofibrils, do not generate a heartbeat, and die around the time of hatching (Humphrey 1972;Lemanski 1973). This mutation has been attributed to defects in the expression of tropomyosin (Zajdel et al 1999), troponin (Sferrazza et al 2007;Zhang et al 2007), and noncoding RNA (Lemanski et al 1996;Zhang et al 2003;Kochegarov et al 2013), and has been associated with a point mutation in the noncoding myofibril-inducing RNA (MIR) (Zhang et al 2003). Our analyses provide strong evidence that the genetic basis of the mutation lies in a large deletion that resulted in the loss of internal tnnt2 exons.…”

Section: Tnnt2 and Cardiacmentioning

confidence: 99%

A chromosome-scale assembly of the axolotl genome

et al. 2019

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The axolotl (Ambystoma mexicanum) provides critical models for studying regeneration, evolution, and development. However, its large genome (∼32 Gb) presents a formidable barrier to genetic analyses. Recent efforts have yielded genome assemblies consisting of thousands of unordered scaffolds that resolve gene structures, but do not yet permit large-scale analyses of genome structure and function. We adapted an established mapping approach to leverage dense SNP typing information and for the first time assemble the axolotl genome into 14 chromosomes. Moreover, we used fluorescence in situ hybridization to verify the structure of these 14 scaffolds and assign each to its corresponding physical chromosome. This new assembly covers 27.3 Gb and encompasses 94% of annotated gene models on chromosomal scaffolds. We show the assembly's utility by resolving genome-wide orthologies between the axolotl and other vertebrates, identifying the footprints of historical introgression events that occurred during the development of axolotl genetic stocks, and precisely mapping several phenotypes including a large deletion underlying the cardiac mutant. This chromosome-scale assembly will greatly facilitate studies of the axolotl in biological research.

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Role of myofibril-inducing RNA in cardiac TnT expression in developing Mexican axolotl

Cited by 6 publications

References 20 publications

Myofibril-Inducing RNA (MIR) is essential for tropomyosin expression and myofibrillogenesis in axolotl hearts

Myofibril-Inducing RNA (MIR) is essential for tropomyosin expression and myofibrillogenesis in axolotl hearts

Identification of a truncated form of methionine sulfoxide reductase a expressed in mouse embryonic stem cells

A chromosome-scale assembly of the axolotl genome

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