The phosphorylation and transcriptional competence of the free cytoplasmic form and the virion form of NS protein of vesicular stomatitis virus (VSV-Indiana/Mudd-Summers) were compared. NS protein is known to exist in two distinct phosphorylated states, NS1 and NS2, that are resolvable by gel electrophoresis. In vitro phosphorylation of virion NS protein by the viral L protein-associated protein kinase resulted in the phosphorylation of both NS1 and NS2. However, in the presence of the N-RNA complex, the NS2 form was preferentially phosphorylated. A cellular protein kinase activity, found in cytoplasmic extracts from VSV-infected or uninfected cells, preferentially phosphorylated NS1, which did not undergo dephosphorylation by cellular phosphatase and also did not convert to NS2. In contrast, the virion or cellular NS2 which had been phosphorylated in vivo or in vitro could be rapidly dephosphorylated by a cellular phosphatase. Cytoplasmic NS protein was found to be fully capable of binding to the virion N-RNA template, and in conjunction with L protein, it participated in synthesis of the leader RNA and five mRNA species of VSV. Moreover, under these conditions, neither cellular phosphatase nor cellular ribonuclease was able to bind to reconstituted nucleocapsids. Binding of cytoplasmic NS to the virion N-RNA template in the presence of L protein resulted in a large and preferential enhancement of NS2 phosphorylation. A protein kinase activity, which phosphorylated NS protein in vitro, was found to be associated with the N-RNA template. This activity appeared to be very tightly bound to N-RNA and exhibited absolute specificity for NS protein of the homologous serotype.

Adenosine Triphosphate↗

PubMed1986

Conserved structures among the nucleocapsid proteins of the paramyxoviridae: complete nucleotide sequence of human parainfluenza virus type 3 NP mRNA.

The nucleotide sequence of the mRNA coding for the nucleocapsid protein (NP) of the paramyxovirus, human parainfluenza virus type 3 (PIV-3), has been determined. The NP mRNA was found to contain 1642 bases, excluding poly(A), and encode a protein of 515 amino acids, with a molecular weight of 57,823. Amino acid residues 1 through 420 of PIV-3 NP protein showed extensive sequence homology with the corresponding amino acids of Sendai virus nucleocapsid protein. There was virtually no homology between the last 95 amino acids. Comparison of the NP proteins of PIV-3, Sendai virus, measles virus, and canine distemper virus revealed, from amino acid residues 160 through 390, some conserved areas between the corresponding proteins of these paramyxoviruses. The 5' terminal sequence of PIV-3 NP mRNA (5'-AGGATTAAAG-3') was similar to the conserved sequence (formula; see text) found at the 5' termini of Sendai virus mRNAs. Both PIV-3 NP and Sendai virus mRNAs had a common 3' terminal tetranucleotide (5'-TAAG-3') preceding the poly (A) tail.

Amino Acid Sequence↗

PubMed1986

Complete nucleotide sequence of the matrix protein mRNA of vesicular stomatitis virus (New Jersey serotype).

The complete nucleotide sequence of the mRNA of the matrix (M) protein of vesicular stomatitis virus [New Jersey serotype, VSV(NJ)] was derived from a cDNA clone and mRNA. The mRNA is 758 nucleotides long (excluding polyadenylic acid) and encodes a protein of 229 amino acids. The predicted amino acid sequence was compared with that of the corresponding protein of Indiana serotype [VSV(IND)] and a fish rhabdovirus, spring viremia of carp virus (SVCV). An amino acid identity of 62% was found between the M proteins of VSV(NJ) and VSV(IND) while only 24% was present between VSV(NJ) and SVCV. A highly basic NH2-terminal domain followed by a proline-proline-X-tyrosine sequence was present in all the three M polypeptides. Except for the L gene sequence, the complete nucleotide sequence of the four genes of VSV(NJ) are now known. The comparison of the amino acid sequences between the Indiana and New Jersey serotypes demonstrates a high degree of homology between these genes except for the phosphoprotein gene, NS.

Amino Acid Sequence↗

PubMed1986

Characterization of vaccinia virus transcripts involved in selective inhibition of host protein synthesis.

Vaccinia virus transcripts synthesized in vitro inhibit protein synthesis directed by globin mRNA, EMC RNA, and total cytoplasmic RNA isolated from HeLa and CHO cells in micrococcal nuclease-treated reticulocyte lysates. In contrast, RNA isolated from vaccinia virus-infected cells is efficiently translated in the same system in the presence of the in vitro transcripts (G. Coppola and R. Bablanian (1983), Proc. Natl. Acad. Sci. USA 80, 75-79). In this study, we have fractionated the in vitro transcripts by acid-urea-agarose gel electrophoresis and demonstrated that the smallest RNA fraction inhibited translation of HeLa cell mRNA in vitro but had no effect on vaccinia virus mRNA translation. On the other hand, the larger RNA species inhibited nonselectively. These results indicate that the selective inhibitory activity of vaccinia virus transcripts resides only in the small poly(A)-containing in vitro RNA fraction.

Binding, Competitive↗

PubMed1986

Production of methionine and glutamic acid from n-alkanes by Serratia marcescens var. kiliensis.

A hydrocarbon-utilizing Serratia marcescens var. kiliensis grew and accumulated methionine and glutamic acid in a synthetic medium with hydrocarbon as sole carbon source. n-Hexadecane and ammonium phosphate were found as the most suitable carbon and nitrogen sources, respectively. Optimum pH for growth and methionine production was 7.2, and that for glutamic acid accumulation was 7.4. Yeast extract significantly stimulated growth and amino acid production and could be substituted by cyanocobalamine. Benzylpenicillin, Tween 80, SDS or EDTA did not increase amino acid production. Under optimal cultural conditions in the laboratory the organism produced 1.68 g of glutamic acid and 0.78 g of methionine per litre.

Alkanes↗

PubMed1986

Messenger RNA encoding the phosphoprotein (P) gene of human parainfluenza virus 3 is bicistronic.

The complete nucleotide sequence of the phosphoprotein (P) mRNA of human parainfluenza virus 3 (PIV-3) was derived from two cDNA clones spanning almost the entire P gene. The mRNA, excluding the poly(A) tail, is 2014 nucleotides long and is bicistronic. The first open reading frame (ORF) codes for the phosphoprotein (P) of mol wt 68,860. Seven nucleotides downstream from the first AUG codon, in a +1 reading frame, there is an additional ORF which can code for a polypeptide of mol wt 23,266. The latter protein appears to be similar to the C proteins found in cells infected with several paramyxoviruses. Comparison of the predicted amino acid sequence of the P and C proteins of PIV-3 with the corresponding Sendai virus proteins reveals considerable homology at the C-terminal half. In contrast, the P and C proteins of PIV-3 share very little homology with the measles virus P and C proteins, respectively.

Amino Acid Sequence↗

PubMed1986

Defective interfering particles of VSVNJ (Ogden), generated by heat treatment, contain multiple internal genomic deletions.

Defective interfering (DI) particles have been isolated from a heat-resistant strain of the New Jersey (Ogden) serotype of vesicular stomatitis virus (VSV). Most of these DI particles contain various portions of all five cistrons of VSV. The two largest DI particles, NJ-121 and NJ-PG2, represent approximately 60% of the standard virus genome and contain both the positive and negative strand leader RNA templates. These two DI particles are transcriptionally active and synthesize both the positive and negative strand leader RNAs in vitro. Virion RNA probe-mRNA hybridizations and cDNA probe-virion RNA hybridizations have shown that NJ-121 contains mainly sequences from the L and G genes. In contrast, NJ-PG2 has portions of the sequences from all five genes of VSV. Smaller DI particles, NJ-121a, NJ-121b, NJ-PG1, and NJ-JM2 representing approximately 50, 38, 28, and 25% of the standard virus genome respectively, were also generated. These DI particles did not have sequences complementary to the positive strand leader RNA template. The mRNA hybridization patterns and results of the genomic RNAs hybridizing to cDNAs of N, NS, M, and G genes of these DI particles showed that they contain parts of information from all five cistrons. Most of the DI particles appear to be generated by multiple deletions throughout the standard virus genome. None of these DI particles interfered heterotypically with VSVIND-HR in BHK21, R(B77), or L2 cells. However, they interfered well with infection by VSVNJ (Hazelhurst).

Chromosome Deletion↗

PubMed1986

Carcinoma of prostate and sexual activity.

Explore the source record for details and available documents.

Aged↗

PubMed1986

The management of acute renal failure in intensive care units.

Explore the source record for details and available documents.

Acute Kidney Injury↗

PubMed1986

Primary spinal epidural Ewing's sarcoma.

A rare case of Ewing's sarcoma, originating primarily in the spinal epidural space is reported. Only five similar case reports are available in the English literature. Full recovery from a state of complete paraplegia, due to malignant compression, local recurrence without distant metastases and the longest survival of more than three years and six months, makes a unique combination. Pathology and therapy are briefly discussed and the pertinent literature reviewed.

Adolescent↗