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A L Haenni

Publications and source records attributed to A L Haenni.

At least 73 records · Page 4Linked to original sources

Comparison of three different cell-free systems for turnip yellow mosaic virus RNA translation.

The two proteins of molecular weights 150,000 and 195,000 specific of turnip yellow mosaic virus (TYMV) RNA translation in reticulocyte lysates have now also been detected in two other cell-free systems programmed with TYMV RNA: the wheat germ extract and the Ehrlich ascites cell-free system. The wheat germ extract contains proteases that affect the nascent TYMV polypeptide chains. The specific post-translational maturation of the protein of molecular weight 195,000 known to occur in the reticulocyte lysate has been investigated in the ascites system. An N-terminal fragment and a C-terminal fragment of molecular weights 120,000 and 78,000, respectively, could be detected co-migrating with the post-translational cleavage products observed in the reticulocyte lysate. Similarly, a C-terminally labelled 78,000 molecular weight fragment could be observed in the wheat germ system. The differences between the three in vitro systems with respect to TYMV RNA translation are discussed.

Animals↗

Length requirements for tRNA-specific enzymes and cleavage specificity at the 3' end of turnip yellow mosaic virus RNA.

This paper describes the minimum length of the turnip yellow mosaic virus (TYMV) RNA necessary to fulfill the tRNA-like properties of the viral RNA: 50 to 75 nucleotides and 86 nucleotides from the 3' end of TYMV RNA are sufficient for adenylation and valylation respectively by the Escherichia coli system. The size of the tRNA-like fragments obtained in vitro in the presence of an E. coli, a reticulocyte or a chinese cabbage leaf extract has also been determined. Among the major fragments liberated from the 3' end of TYMV RNA by the three systems are fragments of 117 and 112 nucleotides. In addition, the E. coli extract liberates fragments of 139 and 61 nucleotides, and the reticulocyte lysate fragments of 109, 94, 84, 73 and 46 nucleotides. The cleavage of the viral RNA by several systems in vitro to yield RNA fragments encompassing the tRNA-like sequence suggests that such fragments might also be liberated in vivo.

Base Composition↗

Turnip yellow mosaic virus RNA is aminoacylated in vivo in Chinese cabbage leaves.

Turnip yellow mosaic virus (TYMV) contains a tRNA-like structure as an integral part of its genome. This structure is located at the extreme 3' end of the viral RNA and is the acceptor of valine after 3'-terminal adenylation. It is known that in vitro (with bacterial, yeast, or plant systems) and in vivo (upon microinjection into Xenopus laevis oocytes) a series of tRNA-specific enzymes can recognize this structure in the viral RNA. We report that TYMV RNA is valylated and consequently adenylated in vivo in its natural host, Chinese cabbage leaves. This suggests that the acylated form of the viral RNA could play an important role in the life-cycle of the virus.

Journal Article↗

Proteolytic maturation of the turnip-yellow-mosaic-virus polyprotein coded in vitro occurs by internal catalysis.

The genomic RNA of turnip yellow mosaic virus is translated in vitro into two major high-molecular-weight proteins, the larger of which (Mr 195 000) undergoes post-translational cleavage. The mechanism of formation of the primary cleavage products (Mr 120 000 and Mr 78 000) of the 195 000-Mr protein has been examined. The fact that cleavage partly occurs at a rate insensitive to dilution of the 195 000-Mr protein is suggestive of an intramolecular mechanism of proteolytic maturation.

Genes, Viral↗

Effect of freezing and thawing on the structure of turnip yellow mosaic virus.

The uncoating of turnip yellow mosaic virus in vitro induced by freezing and thawing has been investigated using a variety of biochemical techniques including the aminoacylation capacity of the viral RNA and the ability of the RNA to stimulate protein synthesis, as well as physico-chemical techniques such as sucrose gradient centrifugation and electron microscopy by negative staining. In particular a fluorescence test has been developed that can serve as a routine method to quantify the RNA liberated during the freeze-thaw process. Escape of the viral RNA is a highly cooperative phenomenon: it depends critically on the virus concentration during freezing and thawing. Increasing the ionic strength or including foreign proteins diminish the escape of the RNA. The RNA is not damaged by this treatment and its liberation occurs without disruption of the viral capsid.

Aminoacylation↗

Binding of ribosomes to 5'-terminal leader sequences of eukaryotic messenger RNAs.

Tobacco mosaic virus and turnip yellow mosaic virus genome RNAs, which have long (greater than or equal to 69 nucleotides) 5'-terminal leader sequences preceding the AUG initiation codon, can form disome initiation complexes when incubated with a wheat germ extract and sparsomycin. In tobacco mosaic virus RNA, whereas one ribosome in such complexes presumably occupies the AUG-containing initiation site, the other is located within the RNA leader sequence. The results favor a model of eukaryotic initiation in which ribosomes first bind to the 5' end of the mRNA independently of AUG, then advance to the AUG initiation codon, where protein synthesis ensues.

Base Sequence↗

A restriction map of cauliflower mosaic virus DNA (strain PV 147). Mapping of the cleavage sites of HhaI, SacI, AvaI, PvuII, PstI, XbaI, EcoRI, Bg/II, HincII, HpaII and HindII + III.

The virion-extracted DNA (Mr5 x 10(6)) of cauliflower mosaic virus (CaMV) has three single-stranded interruptions. The mapping of this DNA using eleven restriction endonucleases (HhaI, SacI, AvaI, PvuII, PstI, XbaI, EcoRI, Bg/II, HincII, HpaII and HindII + III) is reported here. The existence of the three single-stranded breaks complicates the identification and the molecular weight determination of fragments produced by HpaII, HindIII and HindII + III. Indeed the electrophoretic mobility of some fragments in which a single-stranded discontinuity is located is modified, and the fluorescence of ethidium bromide complexed with these fragments is reduced as compared to that observed for the other fragments existing in a molar ratio. These drawbacks were overcome by performing experiments of nick-translation of CaMV DNA with Escherichia coli DNA polymerase I. FRom the data it follows that the CaMV DNA molecule bears bears 1 site for HhaI and SacI, 2 for AvaI and PvuII, 3 for PstI, 4 for XbaI, 5 for EcoRI, 6 for Bg/II and HincII, 11 for HpaII and 15 for HindII + III. The corresponding fragments have all been ordered and precisely located providing a suitable map for further investigations connected with the study of the fine structure and the function of the CaMV genome.

DNA Restriction Enzymes↗

Histone mRNA in Xenopus laevis ovaries: identification of the H4 messenger.

RNA extracted from ovaries of adult Xenopus laevis and introduced into a wheat germ cell-free system, directs the synthesis of polypeptides co-electrophoresing with all five histones. By frationation of total ovary RNA through Sephadex G-200 and polyacrylamide gel, RNA was isolated that is translated into histones 4 (H4) only. In addition, this RNA labelled in vitro with [I125] hybridizes to a restriction fragment of cloned sea urchin DNA known to contain the H4 gene.

Animals↗

Studies on the sequence of the 3'-terminal region of turnip-yellow-mosaic-virus RNA.

A fragment representing the 3'-terminal 'tRNA-like' region of turnip yellow mosaic (TYM) virus RNA has been purified following incubation of intact TYM virus RNA with Escherichia coli 'RNase P'. This fragment, which is 112+3-nucleotides long has been completely digested with T1 RNase and pancreatic RNase and all the oligonucleotides present in such digests have been sequenced using 32P-end labelling techniques in vitro. The TYM virus RNA fragment is free of modified nucleosides and does not contain a G-U-U-C-R sequence. Using nuclease P1 from Penicillium citrinum, the sequence of 26 nucleotides from the 5' end and 16 nucleotides from the 3' end of this fragment has been deduced. The nucleotide sequence at the 5' end of the TYM virus RNA fragment indicates that this fragment includes the end of the TYM virus coat protein gene.

Base Sequence↗

In vitro synthesis of turnip yellow mosaic virus coat protein in a wheat germ cell-free system.

Turnip Yellow Mosaic Virus RNA directs the synthesis in vitro of its coat protein in a wheat germ cell-free extract. Optimum conditions for synthesis have been defined, and the effect of spermine on specifically enhancing coat protein formation has been examined. Identity between the in vitro synthesized coat protein and authentic coat protein of Turnip Yellow Mosaic Virus was established by analysis on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, peptide mapping, and immunoprecipitation.

Journal Article↗

tRNA nucleotidyltransferase-catalyzed incorporation of CMP and AMP into RNA-bacteriophage genome fragments.

Fragments of bacteriophage RNAs R17, MS2 and Qbeta obtained by incubation with commercial snake venom phosphodiesterase become substrates of the Escherichia coli tRNA nucleotidyltransferase. The transferase adds back CMP and AMP in conditions in which it remains highly specific of CCA-deprived tRNAs. The results suggest that the fragment from the 3' end of the viral genome and/or possibly one or more internal fragment(s) are recognized by the transferase. These observations might indicate that bacteriophage RNAs contain certain features probably present in all tRNAs and which are recognized by the transferase.

Adenosine Monophosphate↗