Order of Transcription of Genes of Vesicular Stomatitis Virus

• Published in: Proceedings of the National Academy of Sciences - PNAS Vol. 73; no. 2; pp. 442 - 446
• Main Authors: Ball, L. Andrew, White, Carol N.
• Format: Journal Article
• Language: English
• Published: United States National Academy of Sciences of the United States of America 01.02.1976; National Acad Sciences
• Subjects: Cell free system; Dosage; Electrophoresis, Polyacrylamide Gel; Gels; Genes; Genomes; Irradiation; Messenger RNA; Molecular weight; Protein Biosynthesis; Radiation Genetics; RNA; RNA, Messenger - biosynthesis; RNA, Viral - biosynthesis; Transcription, Genetic; Ultraviolet Rays; Vesicular stomatitis Indiana virus - metabolism; Vesicular stomatitis Indiana virus - radiation effects; Viral Proteins - analysis; Viral Proteins - biosynthesis; Viruses
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.73.2.442

The effect of ultraviolet radiation on the expression of the genes of vesicular stomatitis virus (VSV) was studied in a cell-free system which executed coupled transcription and translation of the viral genes. In this system, which contained detergent-activated virus and a cytoplasmic extract of mouse L-cells, three of the five viral proteins (N, NS, and M) were synthesized accurately and efficiently; a putative precursor to the viral glycoprotein (G) was also made, but synthesis of the L protein was not detected. In response to irradiation of VSV, the synthesis of each protein in the coupled system displayed a characteristic single-hit inhibition. This permitted calculation of the apparent target-sizes for expression of the different viral genes relative to the entire genome (3.6 to 4.0 × 106daltons) as measured by loss of infectivity. These are: N, 0.55 × 106; NS, 0.83 × 106; M, 1.12 × 106; G, 1.76 × 106daltons of RNA. Only the N protein gene has a target-size which corresponds to that predicted from the molecular weight of its messenger RNA (molecular weight: 0.55 × 106). The target-sizes for the other three genes are two to four times larger than expected, and are not proportional to the molecular weights of their corresponding messenger RNAs (molecular weights: NS, 0.28 × 106; M, 0.28 × 106; G, 0.7 × 106). This polar effect of UV irradiation is inconsistent with independent transcription of each of the genes of VSV. Rather, the target-sizes appear to be cumulative, suggesting that transcription initiates at a single point on VSV RNA and proceeds in the order 3′- N - NS - M - G - (L) - 5′.