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M Perricaudet

Publications and source records attributed to M Perricaudet.

At least 163 records · Page 9Linked to original sources
PubMed1982

Identification of recombinant plasmids containing DNA sequences derived from the 3' end of ovine thyroglobulin mRNA.

This paper describes the cloning in E. coli of several DNA copies of the 3' portion of ovine thyroglobulin mRNA. The cDNA clones were identified by in situ cloning hybridization to 32P-labelled thyroglobulin cDNA and by positive hybridization-translation assays. The longest thyroglobulin cDNA fragment cloned (psTg21A, 1670 bp), was shown by S1 nuclease mapping to be an unaltered copy of the thyroglobulin mRNA. psTg 21A overlapped with another cDNA fragment (psTg 11, 330 bp) containing the poly(dA)-tail and corresponding therefore to the 3' end of the mRNA. When aligned together the two clones represent more than 20% of the thyroglobulin mRNA length. Another cDNA fragment (psTg 15, 350 bp) was identified as an internal portion of the thyroglobulin mRNA sequence.

Animals↗
PubMed1981

The sequence of the 3' non-coding region of the hexon mRNA discloses a novel adenovirus gene.

We report the sequence of a 1164 nucleotide long DNA segment, located between map positions 59.5 and 62.8 on the adenovirus type 2 genome. The sequence comprises the 701 nucleotides long 3' non-coding region of the hexon mRNA as well as several important processing signals. The sequence revealed unexpectedly that the 3' non-coding region of the hexon mRNA contains a 609 nucleotide long uninterrupted translational reading frame following a potential initiator AUG. A late 14S mRNA, corresponding to the open reading frame, could be identified by S1 nuclease mapping and electronmicroscopy. The mRNA shares a poly(A) addition site with the hexon and pVI mRNAs, and carries a leader sequence which is related, and probably identical, to the tripartite leader, found in late adenovirus mRNAs. The junction between the leader and the body of this novel mRNA is located within the coding part of the hexon gene.

Adenoviruses, Human↗
PubMed1980

Structure of two adenovirus type 12 transforming polypeptides and their evolutionary implications.

The human adenoviruses are classified according to their nucleotide sequence homology and their oncogenic potential in rodents. The left-hand end of the genome of the adenovirus types 5 (ad5), 7 (ad7) and 12 (ad12), which are respectively, non-oncogenic (subgroup C), weakly oncogenic (subgroup B) and highly oncogenic (subgroup A), contains all the genetic information needed to induce and maintain the transformed phenotype. This part of the genome contains the early transcription unit designated E1 which is subdivided in two transcription units E1A and E1B. Two spliced mRNAs are transcribed from the E1A region which codes for several phosphorylated polypeptides. These polypeptides play a key role by controlling the expression of the other early transcription units. The major role of region E1A in adenovirus cell transformation might not be activate the true transforming genes of the region E1B. An additional role probably consists of the activation of some cellular genes as a restriction fragment containing this region can immortalize rodent cells in vitro. An important question is why some adenoviruses are oncogenic and others are not. We report here differences in the structures of the E1A polypeptides from ad7 and ad12, compared to ad5, which may partially account for their differing oncogenicity.

Adenoviruses, Human↗
PubMed1980

Construction of restriction enzyme fragment libraries containing DNA from human adenovirus types 2 and 5.

Restriction-fragment libraries containing adenovirus type 2 (Ad2) DNA have been constructed, using the pBR322 plasmid (Bolivar et al., 1977) as a vector. Clones have been isolated which contain all the HindIII fragments of Ad2 DNA except the terminal G- and K-fragments inserted into the HindIII cleavage site of the vector. All the 13 SmaI-fragments of Ad2 DNA were separately inserted into the PstI site of the pBR322 vector after addition of homopolymeric poly(dG) tails to the fragments and poly(dC) tails to the linearized plasmid. Two large fragments of adenovirus type 5 (AD5) DNA, located between map positions 17.0 and 59.5 and between map positions 59.5 and 97.3, respectively, were cloned using bacteriophage lambda as a vector. All clones, which are described in the present report, are available upon request.

Adenoviruses, Human↗
PubMed1980

The gene for polypeptide IX of adenovirus type 2 and its unspliced messenger RNA.

A 606 base pair segment of adenovirus type 2 DNA which includes the gene for polypeptide IX and its flanking regions has been sequenced. Comparison with the sequence of a cDNA copy of the 9S mRNA for polypeptide IX and direct analysis of its capped oligonucleotide show that the gene and its mRNA are colinear. The 9S mRNA has a length of 485 nucleotides, excluding the poly(A) tract and the m7G of the cap, and contains one single open translational reading frame which is sufficiently long to encode polypeptide IX. This reading frame contains three AUG codons, the first of which is used for initiation of protein synthesis. The mRNA has a 5' noncoding region of 24 nucleotides, excluding the m5G of the cap, and encodes a polypeptide which is 139 amino acids long and unusually rich in serine and alanine. The sequence TATATAA is found 25 nucleotides upstream from the cap site. The polyadenylation site has been identified but cannot be located unambiguously because of the presence of two A residues in the DNA sequence at this position. The 3' noncoding region is 41--43 nucleotides long and contains the hexanucleotide sequence AAUAAA. The mRNA for polypeptide IVa2 is encoded adjacent to that for polypeptide IX and is transcribed in the opposite direction. Sequence analysis revealed an overlap of 11--16 nucleotides, depending on the precise location of the polyadenylation sites, between 3' ends of these mRNAs.

Adenoviruses, Human↗
PubMed1980

Two interferon mRNAs in human fibroblasts: in vitro translation and Escherichia coli cloning studies.

Two mRNA species that produce biologically active interferon were isolated from human fibroblasts and studied by size fractionation and cloning in Escherichia coli plasmid pBR322. The major fibroblast interferon (Hu IFN-beta 1) is coded for by the smaller of the two mRNAs, an 11S species, 900 nucleotides long, which in cell-free systems yields a 20,000 Mr protein. The second interferon mRNA species (Hu IFN-beta 2) is 14S, about 1300 nucleotides long, and codes for another protein of 23,000-26,000 Mr. The two interferon mRNAs do not cross-hybridize. Both are induced by poly(rI.rC), but IFN-beta 2 mRNA is induced to about 10% in cells by cycloheximide treatment alone whereas under these conditions IFN-beta 1 is not induced.

Biological Assay↗
PubMed1980

Predicted structure of two adenovirus tumor antigens.

Early adenovirus type 2(Ad2) mRNA sequences have been cloned by using the pBR322 plasmid as a vector. Two clones that include sequences from region E1B were identified and their DNAs were characterized by hybridization, restriction enzyme cleavage, and DNA sequence analysis. The results showed that the clones were derived from two different spliced mRNAs. By combining our results with the established DNA sequence for region E1B of the closely related adenovirus type 5[Maat, J., van Beveren, C.P. & van Ormondt, H. (1980) Gene, in press] it was possible to deduce the structure of a 13S and a 22S mRNA. The two mRNAs differ from each other by the size of their intervening sequences. If translation starts at the first AUG following the cap, the 22S mRNA encodes a Mr 67,000 polypeptide that is terminated by a UGA stop codon located immediately before the splice, whereas the 13S mRNA encodes a Mr 20,000 polypeptide that is translated in different reading frames before and after the splice. The Mr 20,000 and 67,000 polypeptides correspond in molecular weight to two proteins that invariably are precipitated from infected cell extracts by antisera from animals carrying adenovirus-induced tumors.

Adenoviruses, Human↗
PubMed1979

Purification of RNA-DNA hybrids by exclusion chromatography.

A simple method for selection of RNA-DNA hybrids has been developed and applied to the purification of adenovirus-specific messenger RNA. Cytoplasmic RNA prepared from adenovirus type 2 (ad2)-infected HeLa cells or from an ad2-transformed rat cell line was hybridized in solution to the complementary strands of ad2 DNA. The hybridization mixture was subsequently fractionated by chromatography on a Sepharose 2B column. The intact probe DNA as well as the RNA-DNA hybrids are excluded from the gel matrix and elute with the void volume. Nonhybridized RNA, in contrast, is included into the gel matrix and elutes as a broad peak well separated from the excluded fractions. Fractions corresponding to the void volume, were collected and the RNA-DNA hybrids were denatured in 90% formamide. The selected RNA was separated from the DNA by affinity chromatography on poly(U)-Sepharose. Restriction endonuclease fragments of DNA with a large enough size to make them excluded from the agarose column were also used for hybridization. In these experiments hybridizations were carried out under conditions which would allow R-loop formation (Thomas, M., White, R.L., and Davis, R.W. (1976) Proc. Natl. Acad. Sci. U.S.A. 73, 2294-2298) and the hybridized RNA was separated from unhybridized RNA by Sepharose chromatography. The validity of the method was demonstrated by programming an in vitro protein-synthesizing system with selected RNA.

Adenoviruses, Human↗
PubMed1978

Bacteriophage lambda and plasmid vectors, allowing fusion of cloned genes in each of the three translational phases.

We have constructed vectors from bacteriophage lambda and from plasmid pBR322 having a single EcoRI restriction site which is immediately downstream from the lac UV5 promotor. Each vector allows the fusion of a cloned gene to the lac Z gene in a different phase relative to the translation initiation codon of the lac Z gene. These vectors were constructed through modification of the initial EcoRI restriction site by S1 endonuclease treatment and then addition of octadeoxyribonucleotides (EcoRI linkers), which shifted the restriction site by 2 or 4 nucleotides. Used in combination these vectors should allow translation of a cloned gene in any one of the three coding phases. The bacteriophages vectors are certified as B2 (EK2) safety level vectors by the French "recombinaison génétique in vitro" committee (D.G.R.S.T.).

Base Sequence↗
PubMed1976

Propagation in E. coli of bacteriophage lambda with integrated fragments of adenovirus 2 DNA.

Hybrid genomes of bacteriophage lambda with integrated fragments of adenovirus type 2 (Ad2) DNA have been constructed in vitro and propagated in E. coli. DNA from a derivative of bacteriophage lambdaplac5 (Rambach and Tiollais, 1974) was used as a vector. The two fragments of the vector DNA contain all the essential genes for the replication of the lambda DNA but are too short to be encapsidated. Insertion of DNA is therefore essential for plaque formation which constitutes a selection method for phages containing hybrid genomes. Fragments EcoRI-B and EcoRI-F of Ad2 DNA were purified, and separately ligated with the vector fragments. Clones of hybrid phage could readily be isolated. Two clones of hybrid phage containing fragment EcoRI-B inserted in opposite directions were used to study the transcription of adenovirus-specific sequences. Hybridization experiments showed that transcripts from both strands of fragment Ad2-Eco RI-B could be detected and that transcription probably was controlled by the "early" leftward and the "late" rightward promoters on the lambda genome. No polypeptides specified by the adenovirus fragment have so far been identified.

Adenoviridae↗