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Common core sequences are found in skeletal muscle slow- and fast-fiber-type-specific regulatory elements.

ABSTRACT: The molecular mechanisms generating muscle diversity during development are unknown. The phenotypic properties of slow- and fast-twitch myofibers are determined by the selective transcription of genes coding for contractile proteins and metabolic enzymes in these muscles, properties that fail to develop in cultured muscle. Using transgenic mice, we have identified regulatory elements in the evolutionarily related troponin slow (TnIs) and fast (TnIf) genes that confer specific transcription in either slow or fast muscles. Analysis of serial deletions of the rat TnIs upstream region revealed that sequences between kb -0.95 and -0.5 are necessary to confer slow-fiber-specific transcription; the -0.5-kb fragment containing the basal promoter was inactive in five transgenic mouse lines tested. We identified a 128-bp regulatory element residing at kb -0.8 that, when linked to the -0.5-kb TnIs promoter, specifically confers transcription to slow-twitch muscles. To identify sequences directing fast-fiber-specific transcription, we generated transgenic mice harboring a construct containing the TnIs kb -0.5 promoter fused to a 144-bp enhancer derived from the quail TnIf gene. Mice harboring the TnIf/TnIs chimera construct expressed the transgene in fast but not in slow muscles, indicating that these regulatory elements are sufficient to confer fiber-type-specific transcription. Alignment of rat TnIs and quail TnIf regulatory sequences indicates that there is a conserved spatial organization of core elements, namely, an E box, a CCAC box, a MEF-2-like sequence, and a previously uncharacterized motif. The core elements were shown to bind their cognate factors by electrophoretic mobility shift assays, and their mutation demonstrated that the TnIs CCAC and E boxes are necessary for transgene expression. Our results suggest that the interaction of closely related transcriptional protein-DNA complexes is utilized to specify fiber type diversity.

SUBMITTER: Nakayama M

PROVIDER: S-EPMC231230 | biostudies-other | 1996 May

REPOSITORIES: biostudies-other

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Common core sequences are found in skeletal muscle slow- and fast-fiber-type-specific regulatory elements.

Nakayama M M Stauffer J J Cheng J J Banerjee-Basu S S Wawrousek E E Buonanno A A

Molecular and cellular biology 19960501 5

The molecular mechanisms generating muscle diversity during development are unknown. The phenotypic properties of slow- and fast-twitch myofibers are determined by the selective transcription of genes coding for contractile proteins and metabolic enzymes in these muscles, properties that fail to develop in cultured muscle. Using transgenic mice, we have identified regulatory elements in the evolutionarily related troponin slow (TnIs) and fast (TnIf) genes that confer specific transcription in ei ...[more]

PMID: 8628309

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Project description:The contractile properties of fast-twitch and slow-twitch skeletal muscles are primarily determined by the myosin isoform content and modulated by a variety of sarcomere proteins. X-ray diffraction studies of regulatory mechanisms in muscle contraction have focused predominately on fast- or mixed-fibre muscle with slow muscle being much less studied. Here, we used time-resolved X-ray diffraction to investigate the dynamic behaviour of the myofilament proteins in relatively pure slow-twitch-fibre rat soleus (SOL) and pure fast-twitch-fibre rat extensor digitorum longus (EDL) muscle during twitch and tetanic contractions at optimal length. During twitch contractions the diffraction signatures indicating a transition in the myosin heads from ordered OFF states, where heads are held close to the thick filament backbone, to disordered ON states, where heads are free to bind to thin filaments, were found in EDL and not in SOL muscle. During tetanic contraction, changes in the disposition of myosin heads as active tension develops is a quasi-stepwise process in EDL muscle whereas in SOL muscle this relationship appears to be linear. The observed reduced extensibility of the thick filaments in SOL muscle as compared to EDL muscles indicates a molecular basis for this behaviour. These data indicate that for the EDL, thick filament activation is a cooperative strain-induced mechano-sensing mechanism, whereas for the SOL, thick filament activation has a more graded response. These different approaches to thick filament regulation in fast- and slow-twitch muscles may be adaptations for short-duration, strong contractions versus sustained, finely controlled contractions, respectively. KEY POINTS: Fast-twitch muscle and slow-twitch muscle are optimized for strong, short-duration contractions and for tonic postural activity, respectively. Structural events (OFF to ON transitions) in the myosin-containing thick filaments in fast muscle help determine the timing and strength of contractions, but these have not been studied in slow-twitch muscle. The X-ray diffraction signatures of structural OFF to ON transitions are different in fast extensor digitorum longus (EDL) and slow soleus (SOL) muscle, being completely absent during twitches in soleus muscle and blunted during tetanic contractions SOL as compared to EDL Quasi-stepwise thick filament structural OFF to ON transitions in fast twitch muscle may be an adaptation for rapid, ballistic movements, whereas more graded OFF to ON structural transitions in slow-twitch muscle may be an adaptation for slower, finer motions.

Project description:This study evaluated the effects of muscle fiber characteristics on meat quality traits in 45 female fast- and slow-growing ducks. Three duck breeds at typical market ages were selected and slaughtered, including fast-growing ducks (Cherry Valley duck) and slow-growing ducks (Small-sized Beijing duck and Liancheng White duck). M. pectoralis major (PM), m. soleus (SOL), m. gastrocnemius (GAS) and m. extensor digitorum longus (EDL) were used to assess muscle fiber characteristics as well as meat quality properties. The results showed that the fiber compositions in PM, GAS, and EDL muscles only consisted of fast-twitch fibers irrespective of the breeds, while a low percentage of slow-twitch fibers were observed in slow-growing ducks (17.03% and 29.14%). The significant clear differences of fiber diameter, fiber density and fiber cross-sectional area (CSA) was observed among three duck breeds. Small-sized Beijing ducks had the highest diameter and cross-sectional fiber area coupled with a dramatically lowest fiber density when compared to other 2 breeds both in breast and leg muscles. In addition, the meat quality traits such as moisture content, release water, and intramuscular fat content were significantly affected by the breeds. Slow-growing ducks, especially Liancheng White ducks, exhibited higher release water, intramuscular fat content, as well as lower moisture content (P < 0.05) compared to the fast-growing ducks. The lower pH24 h value and shear force tended to be present in breast of Liancheng White ducks (P < 0.05). The higher protein content and collagen content were detected in breast of Liancheng White ducks and the leg muscle of Small-sized Beijing ducks (P < 0.05), respectively. Finally, the correlation coefficients between muscle fiber characteristics and meat quality showed that the diameter, density and CSA of fibers had a moderate or significant correlation with pH, shear force value, moisture content, and protein content of meat in fast-growing ducks. In slow-growing ducks, muscle fiber characteristics had a moderate or significant correlation with pH, shear force value, release water, protein content, and intramuscular fat content of meat. These results indicated that muscle fiber characteristics is a useful parameter to explain in parts the variation of meat quality including pH, shear force value, and protein content of meat, both in slow-growing ducks and fast-growing ducks.

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Common core sequences are found in skeletal muscle slow- and fast-fiber-type-specific regulatory elements.

Publications

Common core sequences are found in skeletal muscle slow- and fast-fiber-type-specific regulatory elements.

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