ABSTRACT
It has been shown by bioinformatic methods that regions of the Bombyx mori viral nuclear polyhedrosis genome encoded two small RNA--snc RNA-1 and snc RNA-2, which could perform a structural function in polyhedra crystals formation. The aim of this work was identification of the nucleotide sequence of small non-coding RNAs, predicted by bioinformatic methods in B. mori polyhedra. The following methods have been used: polymerase chain reaction, agarose gel electrophoresis, the cloning of PCR products, sequencing. There were first determined nucleotide sequences of snc RNA-1 and snc RNA-2 ofpolyhedrin mRNA complementary regions which are included in B. mori polyhedra. These RNAs have 100% identity with bioinformatic predicted sequences. These results confirmed our bioinformatic approach to the search for small RNAs encoded in B. mori nuclear polyhedrosis virus genome.
Subject(s)
Computational Biology , Genome, Viral , Nucleopolyhedroviruses/genetics , RNA, Small Untranslated/chemistry , RNA, Viral/chemistry , Viral Structural Proteins/chemistry , Animals , Base Sequence , Bombyx/virology , Cloning, Molecular , Molecular Sequence Data , Nucleopolyhedroviruses/chemistry , Occlusion Body Matrix Proteins , Polymerase Chain Reaction , RNA, Small Untranslated/genetics , RNA, Viral/genetics , Sequence Analysis, DNA , Viral Structural Proteins/geneticsABSTRACT
By gel-filtration through Sephacryl S-300 it was shown that RNP A complex present in polyhedra of Bombyx mori nuclear polyhedrosis virus has molecular weight (M(w)) about 700 kDa. It was shown that RNP A with M(w) 788 kDa is composed of two polyhedrin 13S-associates with M(w) 342 kDa, two p14 polypeptide with M(w) 14 kDa, two 21 kDa small non-coded RNAs and two 17 kDa small non-coded RNAs. The model of RNP A formation from components making it is proposed. The complex role in the course of polyhedron formation and its role in the course of infection are discussed.
Subject(s)
Bombyx/virology , Nucleopolyhedroviruses/chemistry , RNA, Viral/chemistry , Ribonucleoproteins/chemistry , Viral Structural Proteins/chemistry , Animals , Cell Nucleus/virology , Chromatography, Gel , Cytosol/virology , Larva/virology , Models, Molecular , Nucleopolyhedroviruses/genetics , Nucleopolyhedroviruses/isolation & purification , RNA, Small Untranslated/chemistry , RNA, Small Untranslated/genetics , RNA, Viral/genetics , Ribonucleoproteins/genetics , Viral Structural Proteins/geneticsABSTRACT
It was shown using bioinformatic approach by analysis of alternative transcriptes secondary structure, that A. californica nuclear polyhedrosis virus gene ph encoded two mature miRNAs and three potential miRNAs. Gene orf1629 complementary to gene ph did not encode miRNAs and pre-miRNA-Cs. Gene p10 encodes mature and potential miRNA. Gene p74 located on complementary chain encodes three mature miRNAs.
Subject(s)
Genes, Viral , MicroRNAs , Nucleopolyhedroviruses , Promoter Regions, Genetic , RNA, Viral , Transcription, Genetic , Algorithms , Animals , DNA, Intergenic/chemistry , Genome, Viral , Inverted Repeat Sequences , MicroRNAs/chemistry , MicroRNAs/genetics , Molecular Sequence Data , Moths/virology , Nucleopolyhedroviruses/chemistry , Nucleopolyhedroviruses/genetics , RNA, Viral/chemistry , RNA, Viral/genetics , SoftwareABSTRACT
The search of miRNA genes in Bombyx mori nuclear polyhedrosis virus genome region complementary to very late genes has been carried out. The search miRNA algorithm in silico was developed by us. It was shown that NPV B. mori genome region containing orf4 gene complementary to ph gene encodes the potential miRNA. NPV B. mori genome region containing p74 gene complementary to p10 gene encodes mature miRNA and potential miRNA. The genome region containing orf1629 encodes two small non-coding RNAs complementary to orf 5'-end of polyhedrin miRNA. From obtained results it is proposed that two small noncoding RNAs complementary to regions of polyhedrin miRNA are included in polyhedra.
Subject(s)
Algorithms , Computational Biology/methods , Genes, Viral , Genome, Viral , MicroRNAs/genetics , Nucleopolyhedroviruses/genetics , Viral Structural Proteins/genetics , Animals , Base Sequence , Bombyx/virology , DNA, Viral/chemistry , DNA, Viral/genetics , Data Mining , Inverted Repeat Sequences , MicroRNAs/chemistry , Molecular Sequence Data , Open Reading Frames , Promoter Regions, Genetic , RNA Folding , Viral Structural Proteins/chemistryABSTRACT
It was shown that two small RNAs about 65 and 55 nucleotides long included in NPV B. mori polyhedra form with polypeptides p29 and p14 specific RNP-complexes with molecular weights of 50 and 31 kDa, respectively. Both complexes form high-molecular weight complex with polyhedrin. Origin and nature of p29 and p14 polypeptides are discussed.
