Abstract
When actinomycin D-treated chick fibroblasts were labeled with 3H-uridine for varying periods during the log phase of Semliki Forest virus infection, radioactivity was found associated with different cytoplasmic fractions. After a 1-min period of labeling, it appeared in a large cytoplasmic structure which was seen in electron micrographs of infected cells. Sediments of sucrose density gradients of cytoplasmic extracts of these cells also contained these structures. Three forms of viral ribonucleic acid (RNA) were associated with this cytoplasmic structure: a ribonuclease-sensitive 42S form identical to the RNA of the mature virus, a ribonuclease-sensitive 26S form, and a ribonuclease-resistant 20S form. After a 5- to 10-min labeling period, radioactivity was associated with a ribonuclease-sensitive 65S cytoplasmic fraction which contained only the 26S RNA form. Finally, after a 1-hr labeling period, a 140S ribonuclease-resistant particle was the most prominent radioactive structure in the cytoplasm. This particle contained only 42S viral RNA. Negative-contrast electron micrographs of the 140S particle and the virion demonstrated structural differences between them. The base compositions of the 42S and 26S viral RNA forms were not significantly different. The base composition of the 20S form differed significantly from that of the other two viral RNA forms, but the values obtained for the mole fractions of the bases present in the 20S form differed, and depended on the period during the virus growth cycle in which 32P was present. These results suggested that viral RNA originated in the large cytoplasmic body. The 20S RNA appeared to be a structure engaged in viral RNA replication and the 140S particle appeared to be a virus precursor.
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Selected References
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- BROWN F., CARTWRIGHT B. VIRUS SPECIFIC RIBONUCLEIC ACIDS IN BABY HAMSTER KIDNEY CELLS INFECTED WITH FOOT-AND-MOUTH DISEASE VIRUS. Nature. 1964 Nov 28;204:855–856. doi: 10.1038/204855a0. [DOI] [PubMed] [Google Scholar]
- Bishop J. M., Summers D. F., Levintow L. Characterization of ribonuclease-resistant RNA from poliovirus-infected HeLa cells. Proc Natl Acad Sci U S A. 1965 Oct;54(4):1273–1281. doi: 10.1073/pnas.54.4.1273. [DOI] [PMC free article] [PubMed] [Google Scholar]
- CHENG P. Y. Purification, size, and morphology of a mosquitoborne animal virus, Semliki Forest virus. Virology. 1961 May;14:124–131. doi: 10.1016/0042-6822(61)90139-8. [DOI] [PubMed] [Google Scholar]
- DALGARNO L., MARTIN E. M. STUDIES ON EMC VIRAL RNA SYNTHESIS AND ITS LOCALIZATION IN INFECTED KREBS ASCITES CELLS. Virology. 1965 Jul;26:450–465. doi: 10.1016/0042-6822(65)90008-5. [DOI] [PubMed] [Google Scholar]
- Dalgarno L., Martin E. M., Liu S. L., Work T. S. Characterization of the products formed by the RNA polymerases of cells infected with encephalomyocarditis virus. J Mol Biol. 1966 Jan;15(1):77–91. doi: 10.1016/s0022-2836(66)80210-3. [DOI] [PubMed] [Google Scholar]
- ERIKSON R. L., FENWICK M. L., FRANKLIN R. M. REPLICATION OF BACTERIOPHAGE RNA: STUDIES ON THE FATE OF PARENTAL RNA. J Mol Biol. 1964 Dec;10:519–529. doi: 10.1016/s0022-2836(64)80070-x. [DOI] [PubMed] [Google Scholar]
- Friedman R. M., Levy H. B., Carter W. B. Replication of semliki forest virus: three forms of viral RNA produced during infection. Proc Natl Acad Sci U S A. 1966 Aug;56(2):440–446. doi: 10.1073/pnas.56.2.440. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Friedman R. M., Sonnabend J. A. Inhibition of interferon action by puromycin. J Immunol. 1965 Oct;95(4):696–703. [PubMed] [Google Scholar]
- Girard M., Baltimore D. The effect of HeLa cell cytoplasm on the rate of sedimentation of RNA. Proc Natl Acad Sci U S A. 1966 Sep;56(3):999–1002. doi: 10.1073/pnas.56.3.999. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Joklik W. K., Becker Y. Studies on the genesis of polyribosomes. II. The association of nascent messenger RNA with the 40 S subribosomal particle. J Mol Biol. 1965 Sep;13(2):511–520. doi: 10.1016/s0022-2836(65)80113-9. [DOI] [PubMed] [Google Scholar]
- KELLY R. B., GOULD J. L., SINSHEIMER R. L. THE REPLICATION OF BACTERIOPHAGE MS2. IV. RNA COMPONENTS SPECIFICALLY ASSOCIATED WITH INFECTION. J Mol Biol. 1965 Mar;11:562–575. doi: 10.1016/s0022-2836(65)80011-0. [DOI] [PubMed] [Google Scholar]
- LEVINTOW L. THE BIOCHEMISTRY OF VIRUS REPLICATION. Annu Rev Biochem. 1965;34:487–526. doi: 10.1146/annurev.bi.34.070165.002415. [DOI] [PubMed] [Google Scholar]
- LWOFF A., HORNE R., TOURNIER P. A system of viruses. Cold Spring Harb Symp Quant Biol. 1962;27:51–55. doi: 10.1101/sqb.1962.027.001.008. [DOI] [PubMed] [Google Scholar]
- MARTIN R. G., AMES B. N. A method for determining the sedimentation behavior of enzymes: application to protein mixtures. J Biol Chem. 1961 May;236:1372–1379. [PubMed] [Google Scholar]
- MORGAN C., HOWE C., ROSE H. M. Structure and development of viruses as observed in the electron microscope. V. Western equine encephalomyelitis virus. J Exp Med. 1961 Jan 1;113:219–234. doi: 10.1084/jem.113.1.219. [DOI] [PMC free article] [PubMed] [Google Scholar]
- MUSSGAY M. GROWTH CYCLE OF ARBOVIRUSES IN VERTEBRATE AND ARTHROPOD CELLS. Prog Med Virol. 1964;6:193–267. [PubMed] [Google Scholar]
- MUSSGAY M., WEIBEL J. Electron microscopic and biological studies on the growth of Venezuelan equine encephalitis virus in KB cells. Virology. 1962 Jan;16:52–62. doi: 10.1016/0042-6822(62)90201-5. [DOI] [PubMed] [Google Scholar]
- Mécs E., Sonnabend J. A., Martin E. M., Fantes K. H. The effect of interferon on the synthesis of RNA in chick cells infected with Semliki forest virus. J Gen Virol. 1967 Jan;1(1):25–40. doi: 10.1099/0022-1317-1-1-25. [DOI] [PubMed] [Google Scholar]
- OCHOA S., WEISSMANN C., BORST P., BURDON R. H., BILLETER M. A. REPLICATION OF VIRAL RNA. Fed Proc. 1964 Nov-Dec;23:1285–1296. [PubMed] [Google Scholar]
- Osterrieth P. M., Calberg-Bacq C. M. Changes in morphology, infectivity and haemagglutinating activity of Semliki Forest virus produced by the treatment with caseinase C from Streptomyces albus G. J Gen Microbiol. 1966 Apr;43(1):19–30. doi: 10.1099/00221287-43-1-19. [DOI] [PubMed] [Google Scholar]
- PENMAN S., BECKER Y., DARNELL J. E. A CYTOPLASMIC STRUCTURE INVOLVED IN THE SYNTHESIS AND ASSEMBLY OF POLIOVIRUS COMPONENTS. J Mol Biol. 1964 Apr;8:541–555. doi: 10.1016/s0022-2836(64)80010-3. [DOI] [PubMed] [Google Scholar]
- PFEFFERKORN E. R., HUNTER H. S. PURIFICATION AND PARTIAL CHEMICAL ANALYSIS OF SINDBIS VIRUS. Virology. 1963 Jul;20:433–445. doi: 10.1016/0042-6822(63)90092-8. [DOI] [PubMed] [Google Scholar]
- Penman S., Scherrer K., Becker Y., Darnell J. E. POLYRIBOSOMES IN NORMAL AND POLIOVIRUS-INFECTED HELA CELLS AND THEIR RELATIONSHIP TO MESSENGER-RNA. Proc Natl Acad Sci U S A. 1963 May;49(5):654–662. doi: 10.1073/pnas.49.5.654. [DOI] [PMC free article] [PubMed] [Google Scholar]
- SEBRING E. D., SALZMAN N. P. AN IMPROVED PROCEDURE FOR MEASURING THE DISTRIBUTION OF P32O4--AMONG THE NUCLEOTIDES OF RIBONUCLEIC ACID. Anal Biochem. 1964 May;8:126–129. doi: 10.1016/0003-2697(64)90177-0. [DOI] [PubMed] [Google Scholar]
- TAYLOR J. STUDIES ON THE MECHANISM OF ACTION OF INTERFERON. I. INTERFERON ACTION AND RNA SYNTHESIS IN CHICK EMBRYO FIBROBLASTS INFECTED WITH SEMLIKI FOREST VIRUS. Virology. 1965 Mar;25:340–349. doi: 10.1016/0042-6822(65)90053-x. [DOI] [PubMed] [Google Scholar]