Messenger RNA for myosin polypeptides: Isolation from single myogenic cell cultures

Strohman, Richard C.; Moss, Paul S.; Micou-Eastwood, Julie; Spector, Dennis; Przybyla, Alan; Paterson, Bruce M.

doi:10.1016/0092-8674(77)90220-3

Cited by 410 publications

(151 citation statements)

References 23 publications

Supporting

Mentioning

150

Contrasting

Order By: Relevance

“…RNA Blotting and In Vitro Nuclear Run-on Transcription Assays RNA (Strohman et al, 1977) and nuclei (Gilley et al, 1990) were isolated at various times from 12-20 liters of P. tetraurelia, growing in log phase, after deciliation (Machemer and Ogura, 1979). Transcriptional activity was measured using the run-on assay (Gilley et al, 1990).…”

Section: Genomic Library Screening and Dna Sequencingmentioning

confidence: 99%

The dynein genes of Paramecium tetraurelia: the structure and expression of the ciliary beta and cytoplasmic heavy chains.

Kandl

Forney

Asai

1995

MBoC

View full text Add to dashboard Cite

The genes encoding two Paramecium dynein heavy chains, DHC-6 and DHC-8, have been cloned and sequenced. Sequence-specific antibodies demonstrate that DHC-6 encodes ciliary outer arm beta-chain and DHC-8 encodes a cytoplasmic dynein heavy chain. Therefore, this study is the first opportunity to compare the primary structures and expression of two heavy chains representing the two functional classes of dynein expressed in the same cell. Deciliation of paramecia results in the accumulation of mRNA from DHC-6, but not DHC-8. Nuclear run-on transcription experiments demonstrate that this increase in the steady state concentration of DHC-6 mRNA is a consequence of a rapid induction of transcription in response to deciliation. This is the first demonstration that dynein, like other axonemal components, is transcriptionally regulated during reciliation. Analyses of the sequences of the two Paramecium dyneins and the dynein heavy chains from other organisms indicate that the heavy chain can be divided into three regions: 1) the sequence of the central catalytic domain is conserved among all dyneins; 2) the tail domain sequence, consisting of the N-terminal 1200 residues, differentiates between axonemal and cytoplasmic dyneins; and 3) the N-terminal 200 residues are the most divergent and appear to classify the isoforms. The organization of the heavy chain predicts that the variable tail domain may be sufficient to target the dynein to the appropriate place in the cell.

show abstract

Section: Genomic Library Screening and Dna Sequencingmentioning

confidence: 99%

The dynein genes of Paramecium tetraurelia: the structure and expression of the ciliary beta and cytoplasmic heavy chains.

Kandl

Forney

Asai

1995

MBoC

View full text Add to dashboard Cite

show abstract

“…Total RNA was isolated from cultured cells and from isolated muscle tissue by standard methods (Strohman et al, 1977;Chomczynski and Sacchi, 1987). Gel electrophoresis, blotting, hybridization to specific probes, and densitometric analysis were all performed as described previously (Wade et al, 1990;Sutherland et al, 1991).…”

Section: Rna Isolation and Northern Analysismentioning

confidence: 99%

Identification of a program of contractile protein gene expression initiated upon skeletal muscle differentiation

Sutherland

Esser

Elsom

et al. 1993

Developmental Dynamics

View full text Add to dashboard Cite

The functional diversity of skeletal muscle is largely determined by the combinations of contractile protein isoforms that are expressed in different fibers. Just how the developmental expression of this large array of genes is regulated to give functional phenotypes is thus of great interest. In the present study, we perform a comprehensive analysis of contractile protein isoform mRNA profiles in skeletal muscle systems representing each generation of fiber formed: primary, secondary, and regenerating fibers. We find that in each system examined there is a common pattern of isoform gene expression during early differentiation for 5 of the 6 gene families we have investigated: myosin light chain (MLCI1, MLC2, tropomyosin, troponin (Tn)C, and TnI. We suggest that the common isoform patterns observed together represent a genetic program of skeletal muscle differentiation that is independent of the mature fiber phenotype and is found in all newly formed myotubes. Within each of these contractile protein gene families the program is independent of the isoforms of myosin heavy chain (MHC) expressed. The maintenance of such a program may reflect a specific requirement of the initial differentiation process. 0 1993 Wiley-Liss, Inc.

show abstract

“…The TBS was removed and the cells were dissolved in 8 M-guanidine hydrochloride, 0.1 M-potassium acetate, pH 5.2 (1 ml per plate). The RNA was then purified essentially according to the techniques of Strohman et al (1977). The poly(A)-containing RNA [poly(A) ÷ RNA] was isolated by oligodeoxythymidylate-cellulose chromatography (Aviv & Leder, 1972).…”

Section: Cells and Virusesmentioning

confidence: 99%

“…This concentration of actinomycin D was sufficient to inhibit the transcription of the bulk cellular poly(A) ÷ RNA by greater than 95% (data not shown). The total cellular RNA was isolated by the guanidine hydrochloride technique (Strohman et al, 1977) and the poly(A) + and poly(A)-RNA fractions were separated by oligo(dT)-cellulose chromatography. The results of this analysis are given in Fig.…”

Section: Decay Of Cellular and B77 Asv Rna Species After The Additionmentioning

confidence: 99%

Stabilities of Avian Sarcoma Virus RNAs: Comparison of Subgenomic and Genomic Species with Cellular mRNAs

Dimock

Horikami

et al. 1983

Journal of General Virology

View full text Add to dashboard Cite

SUMMARYThe stabilities of B77 avian sarcoma virus intracellular RNAs were compared to the stability of the total cellular poly(A)-containing RNA by labelling infected chicken embryo fibroblasts with [3H]uridine for 15 h, adding actinomycin D (1 ~tg per ml) to block further transcription of viral RNA, and selecting virus-specific RNA from the total cellular poly(A)-containing RNA at 3 hourly intervals. The three virus-specific RNA species (9-3, 3.3 and 5.4 kilobases) decayed with half-lives of 7-5, 10, and 15 h, respectively, whereas the bulk of the cellular mRNA decayed with a half-life of 13 h. To correlate these decay rates with the disappearance of mRNA activities, the actinomycin D-treated cells were pulse-labelled with [3H]leucine at 3 hourly intervals after the addition of the drug and virus-specific protein synthesis was assayed by immunoprecipitation. The mRNA activity for the precursor to the non-glycosylated viral structural proteins (Pr76g ag) decayed with a half-life of approximately 6 h, whereas the mRNA activity coding for the precursor to the envelope proteins (gPr92 env) decayed with a half-life of 14 h. Thus, the rate of decay of the individual mRNA species corresponded reasonably well with the decay rate for the synthesis of two of the corresponding gene products. The results indicated that the 5-4 kb env mRNA is more stable under these conditions than the 9.3 kb gag mRNA but was not significantly more stable than the bulk of the cellular mRNA. Virus particle production following the addition of actinomycin D was determined by the reverse transcriptase assay and by the incorporation of viral genomic 70S RNA into extracellular virions. Both assays yielded similar results and indicated that particle production was inhibited at a rate (t~ = 4 h) somewhat faster than the decay of Pr769ag synthesis or the disappearance of 9.3 kb RNA. It was established by two independent methods (pulse and chase, and approach to isotope equilibrium), however, that the intracellular halflife of the RNA that is packaged into virions is 6 to 7 h. Tt~us, these results suggest that a single metabolic pool of 9.3 kb RNA exists in avian sarcoma virus-infected cells and is used both as mRNA and as genome RNA.

show abstract

Messenger RNA for myosin polypeptides: Isolation from single myogenic cell cultures

Cited by 410 publications

References 23 publications

The dynein genes of Paramecium tetraurelia: the structure and expression of the ciliary beta and cytoplasmic heavy chains.

The dynein genes of Paramecium tetraurelia: the structure and expression of the ciliary beta and cytoplasmic heavy chains.

Identification of a program of contractile protein gene expression initiated upon skeletal muscle differentiation

Stabilities of Avian Sarcoma Virus RNAs: Comparison of Subgenomic and Genomic Species with Cellular mRNAs

Contact Info

Product

Resources

About