Mapping of Early and Late Transcripts Encoded by the Autographa californica Nuclear Polyhedrosis Virus Genome: Is Viral RNA Spliced?

Abstract

Early and late transcripts were mapped on the Autographa californica nuclear polyhedrosis virus genome by Northern blotting and hybridization with the cloned viral EcoRI fragments. At least 11 early and about 90 late RNAs were compared with over 32 polypeptides synthesized by in vitro translation of hybrid-selected RNA. The latter method, of course, had its limitations also and did not guarantee that all viral RNAs could be detected in this way. A comparison of cytoplasmic and total cellular RNAs showed no clear-cut differences in their size distributions. We found that there were more RNA classes than corresponding proteins encoded by them and mapped by in vitro translation. By using the Berk-Sharp method and analyses of DNA-RNA hybrids by one-dimensional or two-dimensional neutral and alkaline gel electrophoreses, we were unable to adduce evidence for RNA splicing in this viral system. Minor splices, particularly at sites close to the termini of RNA molecules, could not be excluded.

Divergent transcription of early 35- and 94-kilodalton protein genes encoded by the HindIII K genome fragment of the baculovirus Autographa californica nuclear polyhedrosis virus

“…AcNPV transcription is characterized by the synthesis of a multitude of overlapping, polyadenylated RNAs. Transcripts with maximum synthesis during each of the temporal phases have been mapped throughout the viral genome (3,6,20,30). Overlapping nests of transcripts composed of RNAs with identical 5' or identical 3' termini are common (6,21,28), yet splicing does not appear to be involved in the generation of these RNAs (20).…”

Adenovirus types 2 and 5 functions elicit replication and late expression of adenovirus type 12 DNA in hamster cells

“…The total nuclear DNA was extracted by modified standard procedures (43). In some of the experiments, the total RNA (2) or the cytoplasmic RNA (35) was extracted and analyzed for Adl2-specific RNA sequences (24,25).…”

Proteoma do baculovírus Anticarsia gemmatalis múltiplo nucleopoliedrovírus em linhagens celulares distintas e comparação da proteína de envelope GP64 em variantes geográficos.

“…These results are in line with the possibility that tandem transcripts from ROTs in baculovirus are not necessarily translated at once [24][25][26]. Moreover, long mRNAs of several genes have been observed late in infection [27,28,31,63].…”

“…Our preliminary, unpublished data on 367 RNA suppression subtractive hybridization (SSH) showed that most of these cellular 368 proteins had their mRNA level altered during AgMNPV-2D infection [33]. 369 Importantly, these virion-associated host proteins have been identified previously in 370 other enveloped viruses (Vaccinia virus (VV) [63], Herperviruses (AIHV-1 [64], HSV 371 [65] and KSHV [66]), Epstein-Barr virus (EBV) [67], human cytomegalovirus 372 (HCMV) [68], respiratory syncytial virus (RSV) [69], influenza virus (IV) [70] and 373 HIV-1 [71]) and some have already been identified in other baculovirus [22,26], like: 374 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 By using different proteomic methodologies we identified the structural proteins of 380 the AgMNPV-2D isolate. Our complete proteomic studies and immunoblotting 381 experiments confirmed 44 and 33 proteins present in the ODV and BV, respectively 382 ( Fig.…”

“…490 491 Host associated proteins in both phenotypes. Most of the host proteins 492 associated with the AgMNPV-2D were previously reported in the virion structure of 493 other enveloped viruses such as, Vaccinia virus (VV) [63], Herperviruses (AIHV-1 494 [64], HSV [65] and KSHV [66]), Epstein-Barr virus (EBV) [67], human 495 cytomegalovirus (HCMV) [68], respiratory syncytial virus (RSV) [69], influenza 496 virus (IV) [70] and HIV-1 [71]. These were studies carried out with several 497 mammalian viruses infecting mammalian cell lines.…”