Search PubMed⌕ Search

Biomedical subjects

M Linial

Publications and source records attributed to M Linial.

At least 73 records · Page 4Linked to original sources
PubMed1987

MC29 deletion mutants which fail to transform chicken macrophages are competent for transformation of quail macrophages.

A number of MC29 mutants with deleted myc genes have been previously characterized. Many of these mutants have been found to be defective for transformation of chicken macrophages in vitro and for tumor induction in chickens. Such mutants are capable of transforming Japanese quail macrophages in vitro and inducing a high incidence of tumors in Japanese quail. Thus, Japanese quail may contain a factor(s) capable of complementing the defective transforming proteins encoded by some deleted v-myc genes.

Animals↗
PubMed1986

A nicking enzyme from trypanosomatids which specifically affects the topological linking of duplex DNA circles. Purification and characterization.

Newly replicated duplex DNA minicircles of trypanosomal kinetoplast DNA are nicked in both their monomeric and catenated topological states, whereas mature ones are covalently sealed. The possibility that nicking may play a role during kinetoplast DNA replication by affecting the topological interconversions of monomeric DNA minicircles and catenane networks was studied here in vitro using Crithidia fasciculata DNA topoisomerase. An enzyme that catalyzes the nicking of duplex DNA circles has been purified to apparent homogeneity from C. fasciculata cell extracts. The native enzyme has a sedimentation coefficient of 6.8 S and was found to be a dimer with a protomer Mr = 60,000. Nicking of kinetoplast DNA networks by the purified enzyme inhibits their decatenation by the Crithidia DNA topoisomerase but has no effect on the catenation of monomeric DNA minicircles into networks. This differential effect on decatenation versus catenation is specific to the purified nicking enzyme. Random nicking of interlocked DNA minicircles has no detectable effect on the reversibility of the topological reaction. The potential role of Crithidia nicking enzyme in the replication of kinetoplast DNA networks in trypanosomatids is discussed.

Crithidia↗
PubMed1985

Enhanced transcription of c-myc in bursal lymphoma cells requires continuous protein synthesis.

In several bursal lymphoma cell lines in which c-myc transcription is regulated by avian leukosis virus (ALV) long terminal repeat (LTR) sequences, protein synthesis inhibition decreases the transcriptional activity of c-myc as well as other LTR driven viral genes. This decrease in transcription is associated with a change in the chromatin structure of c-myc, as measured by deoxyribonuclease I (DNase I) hypersensitivity, and a shift of transcription from the LTR to the normal c-myc promoter. In contrast, cycloheximide had little or no effect on the transcription of LTR driven genes in infected chicken embryo fibroblasts treated with the drug. These results suggest that a labile, cell type-specific protein may interact with the retroviral LTR and regulate transcription of genes under LTR control. Further, the results demonstrate that the increase in intracellular concentration of c-myc RNA induced by cycloheximide treatment of normal cells is the result of stabilization of this message.

Animals↗
PubMed1985

Transcription of three c-myc exons is enhanced in chicken bursal lymphoma cell lines.

The chicken c-myc gene, as defined by its homology to the v-myc gene of MC29 virus, is comprised of two exons. Using the techniques of runoff transcription, primer extension, and S1 nuclease protection, we demonstrate that there is a third c-myc exon of approximately equal to 345 base pairs (bp) located 0.7 kbp upstream of the 5' end of the v-myc homology. This first exon is transcribed and present in myc mRNA in normal chicken cells. We also examined RNA from five cell lines derived from avian leukosis virus-induced bursal lymphomas. In all these lines, the level of transcription of the 2.2- to 2.5-kbp myc mRNA is increased 30- to 60-fold over normal cells. The myc mRNA in four of these lines also contains increased levels of the first noncoding exon, and evidence is presented that the long terminal repeat (LTR) in the vicinity of c-myc is functioning as an enhancer of c-myc transcription rather than as a promoter in several of these cell lines. In two cell lines in which the viral LTR has integrated between the first and second exons in the proper orientation for downstream promotion of myc, the LTR does not exhibit promoter function. The pattern of c-myc transcription observed by others in a vast majority of avian leukosis virus-induced neoplasms is not observed in any of the five cell lines examined.

Animals↗
PubMed1984

Nucleotide sequence 5' of the chicken c-myc coding region: localization of a noncoding exon that is absent from myc transcripts in most avian leukosis virus-induced lymphomas.

We have determined the nucleotide sequence of the 2.2-kilobase-pair region upstream of the chicken c-myc coding exons. Using RNA blot analysis, we have localized a noncoding exon to a region that is separated from the c-myc coding sequences by an intron of 700-800 base pairs. In most avian leukosis virus-induced lymphomas proviral integration has occurred within, or downstream of, the first exon, thus presumably displacing the regulatory sequences that normally control c-myc expression. More than 70% of the integration sites were clustered in a 250-base-pair region in the first intron, immediately preceding the coding sequences. Sequences from the upstream noncoding exon were absent from the myc transcripts in these lymphomas; RNA transcripts from the normal c-myc allele were not expressed at detectable levels.

Animals↗
PubMed1983

Expression of c-myc RNA in bursal lymphoma cell lines: identification of c-myc-encoded proteins by hybrid-selected translation.

We examined expression of the c-myc locus in four cell lines established from bursal lymphomas induced by avian leukosis virus. In all four lines the level of myc-related RNA was elevated. In three lines a majority of the myc-containing RNAs lacked viral-LTR-related sequences, in contrast to results obtained with primary tumors. This suggests that LTR sequences are not required for maintenance of high level c-myc expression. One line, RP9, has a complex pattern of myc RNAs containing LTR sequences, and one of these RNAs is packaged into virions. Using hybrid selection of RNAs with myc DNA, followed by in vitro translation, we detected translation of myc-related proteins from RNA of all four cell lines. The sizes of these proteins differ among the cell lines. The major polypeptides detected were 64, 57, and 54 kilodaltons. Events leading to elevation of c-myc transcription may be accompanied by alterations in mRNA initiation or processing that generate different protein products.

Animals↗
PubMed1983

RNA and protein encoded by MH2 virus: evidence for subgenomic expression of v-myc.

MH2 and MC29 are highly related myc-containing avian retroviruses. We found that MH2, unlike MC29, synthesizes a 2.6-kilobase subgenomic mRNA containing myc sequences as well as sequences from the 5' end of the genome. A 57-kilodalton protein containing myc, but not gag, sequences (p57myc) was detected by hybrid selection and in vitro translation of RNA from MH2-transformed cells. Gradient separation of MH2 intracellular RNAs indicated that p57myc is encoded by the subgenomic RNA. A highly oncogenic MH2 virus variant (MH2YS3) (M. Linial, Virology 119:382-391, 1982) was shown to encode only p57myc and not P100, the previously described MH2-encoded polyprotein (Hu et al., Virology, 89:162-178, 1978). Cells transformed by subclones of this virus synthesized predominantly the 2.6-kilobase RNA rather than genomic 5.4-kilobase RNA. These results suggest that only p57myc is required for maintenance of the transformed state after MH2 infection.

Animals↗
PubMed1980

Synthesis and processing of viral glycoproteins in two nonconditional mutants of Rous sarcoma virus.

We have studied the pattern of glycoprotein synthesis in two nonconditional mutants of Rous sarcoma virus. One mutant, SE33, produces no viral particles but synthesizes Pr92env, which is cleaved intracellularly to mature glycoproteins. The second mutant, SE521, encodes a gPr92env which is not cleaved to gp85 or gp37 and therefore produces virions with the phenotype of Bryan RSV(-) or NY8. Neither of these mutants have detectable genomic deletions. The study of these mutants has led to the following conclusions. (i) In the absence of particle production or p15 synthesis, gPr92env can be cleaved to the mature glycoprotein which is found on the cell surface. (ii) Noncleaved gPr92env is not packaged into virions but is found on the cell surface. (iii) gPr92env alone can account for subgroup specific viral interference. (iv) gPr92env is probably transported to the cell surface before additional glycosylation or cleavage to mature virion glycoprotein. The nonprocessed precursor of SE521 appears to be glycosylated normally, and thus far we have been unable to determine the basis for the defect in this mutant.

Avian Sarcoma Viruses↗
PubMed1980

Synthesis and processing of polymerase proteins of wild-type and mutant avian retroviruses.

We have studied the biosynthesis of avian retrovirus proteins related to reverse transcriptase in permissive avian embryonic cells. Analysis of immune precipitates from avian sarcoma virus (ASV)-infected cells demonstrated the presence of the 180,000-dalton gag-pol "read-through" protein (Pr180gag-pol) and a 130,000-dalton polypeptide (Pr130gag-pol). Pr130gag-pol was found, in serological and peptide mapping studies, to consist primarily of sequences related to reverse transcriptase and the gag-encoded protein p15. Pr180gag-pol was found to be phosphorylated, whereas Pr130gag-pol was not. In addition, only Pr180gag-pol but not Pr130gag-pol was susceptible to cleavage with the virion protease p15. Although the structure of Pr130gag-pol would suggest that it is generated by removal of a portion of the gag region from Pr180gag-pol, an analysis of labeling kinetics has failed to demonstrate unequivocally whether Pr130gag-pol is a cleavage product of Pr180gag-pol or a primary translation product. We were repeatedly unable to detect either Pr180gag-pol or Pr130gag-pol in virus particles released from the cell, whereas both beta and alpha subunits were readily observed. Several presumed intermediates between Pr130gag-pol and the beta subunit of reverse transcriptase were also observed in virions. These studies indicate cleavage of polyemrase precursors at the time of virus budding. On the basis of these data, we present a processing scheme for the generation of reverse transcriptase subunits. We have also examined reverse transcriptase biosynthesis in cells producing two mutants that fail to package the enzyme. Previous work showed that integrated proviruses of both mutants are missing DNA sequences in pol: one mutant, PH9 (Mason et al., J. Virol. 30:132-140, 1979), contains a deletion near the 3' end of pol, whereas the other, SE52d (linial et al., Virology 87:130-141, 1978), may have inserted a host cell sequence near the 5' end of pol. Neither mutant synthesized Pr180gag-pol or Pr130gag-pol, but instead produced novel proteins comprised of sequences shared with gag proteins plus a region antigenically related to reverse transcriptase. Both proteins were defective as precursors to reverse transcriptase. Whereas Pr180gag-pol and Pr130gag-pol were precipitated by an antiserum raised against p32 (a virion protein derived from the portion of the beta subunit removed during processing of beta to alpha [Schiff and Grandgenett, J. Virol. 28:279-291, 1978]), the novel protein synthesized by PH9 ws not precipitated. This suggets that the alpha subunit is generated by a COOH-terminal cleavage of the beta subunit.

Alpharetrovirus↗
PubMed1980

Avian oncovirus mutant (SE21Q1b) deficient in genomic RNA: characterization of a deletion in the provirus.

We have previously described a nonconditional mutant of avian sarcoma virus (SE21Q1b) which fails to package viral RNA (Gallis et al., Virology 94:146-161, 1979; Linial et al., Cell 15:1371-1381, 1978). Quail cells transformed by SE21Q1b contain normal amounts of intracellular viral mRNA's for src, env, and gag-pol and release particles with the density of normal virus containing a typical complement of virion proteins, including reverse transcriptase. These virions are noninfectious for both chicken and quail cells and contain primarily cellular rather than viral RNA. Analysis by gel electrophoresis of the cellular DNA of quail cells transformed by SE21Q1b after restriction endonuclease digestion indicated the presence of a single provirus. The provirus was located at one site in the genome of the host cell and was flanked by the characteristic terminally repeated sequences derived from the 3' and 5' ends of viral RNA. The only defect detected in the SE21Q1b provirus was a deletion of ca. 150 base pairs of DNA somewhere between 300 and 600 bases from the left (gag-pol) end of the provirus. Analyses of the proviral DNA of cells transformed by wild-type recombinants between SE21Q1b and leukosis viruses reveal that the recombinants no longer contain this deletion. The deletion, therefore, defines a region on the viral RNA which is required for correct packaging of the virion RNA.

Animals↗