Segmented genome and nucleocapsid of La Crosse virus

J. F. Obijeski, D. H L Bishop, E. L. Palmer, F. A. Murphy

Research output: Contribution to journalArticle

79 Citations (Scopus)

Abstract

La Crosse (LAC) virions purified by velocity and equilibrium gradient centrifugation contained three single stranded RNA species. The three segments had sedimentation coefficients of 31S, 25S, and 12S by sodium dodecyl sulfate sucrose gradient centrifugation. By comparison with other viral and cellular RNA species, the LAC viral RNAs had molecular weights of 2.9x106, 1.8x106, and 0.4x106. Phenol sodium dodecyl sulfate extracted LAC virion RNA was not infectious for BHK 21 cell cultures under conditions in which Sindbis viral RNA was infectious. Treatment of LAC virus with the nonionic detergent Triton X 100 and salt released three nucleocapsid structures, each containing one species of virion RNA. The nucleocapsids had sedimentation coefficients of 115S, 90S, and 65S. Negative contrast electron microscopy of the nucleocapsids indicated that they were convoluted, supercoiled, and apparently circular. They had a mean diameter of 10 to 12 nm and modal lengths of 200, 510, and 700 nm (some were even longer). By chemical and enzymatic analysis of purified viral RNA, one type of 5' nucleotide (pppAp) present in the proportion of one per RNA segment was identified. After periodate oxidation, each virion RNA species was labeled by reduction with [3H] sodium borohydride. Taken together, these results suggest that although the nucleocapsids appear as closed loops, the viral RNA has free 5' and 3' ends and is, therefore, not circular.

Original languageEnglish (US)
Pages (from-to)664-675
Number of pages12
JournalJournal of Virology
Volume20
Issue number3
StatePublished - 1976

Fingerprint

La Crosse virus
California encephalitis virus
Nucleocapsid
nucleocapsid
Viral RNA
Virion
Genome
RNA
genome
Centrifugation
virion
Sodium Dodecyl Sulfate
Octoxynol
sodium dodecyl sulfate
Phenol
Detergents
centrifugation
Sucrose
Electron Microscopy
Nucleotides

ASJC Scopus subject areas

  • Immunology

Cite this

Obijeski, J. F., Bishop, D. H. L., Palmer, E. L., & Murphy, F. A. (1976). Segmented genome and nucleocapsid of La Crosse virus. Journal of Virology, 20(3), 664-675.

Segmented genome and nucleocapsid of La Crosse virus. / Obijeski, J. F.; Bishop, D. H L; Palmer, E. L.; Murphy, F. A.

In: Journal of Virology, Vol. 20, No. 3, 1976, p. 664-675.

Research output: Contribution to journalArticle

Obijeski, JF, Bishop, DHL, Palmer, EL & Murphy, FA 1976, 'Segmented genome and nucleocapsid of La Crosse virus', Journal of Virology, vol. 20, no. 3, pp. 664-675.
Obijeski JF, Bishop DHL, Palmer EL, Murphy FA. Segmented genome and nucleocapsid of La Crosse virus. Journal of Virology. 1976;20(3):664-675.
Obijeski, J. F. ; Bishop, D. H L ; Palmer, E. L. ; Murphy, F. A. / Segmented genome and nucleocapsid of La Crosse virus. In: Journal of Virology. 1976 ; Vol. 20, No. 3. pp. 664-675.
@article{7728a1a4aeb14876878f8bd7cde2a0a1,
title = "Segmented genome and nucleocapsid of La Crosse virus",
abstract = "La Crosse (LAC) virions purified by velocity and equilibrium gradient centrifugation contained three single stranded RNA species. The three segments had sedimentation coefficients of 31S, 25S, and 12S by sodium dodecyl sulfate sucrose gradient centrifugation. By comparison with other viral and cellular RNA species, the LAC viral RNAs had molecular weights of 2.9x106, 1.8x106, and 0.4x106. Phenol sodium dodecyl sulfate extracted LAC virion RNA was not infectious for BHK 21 cell cultures under conditions in which Sindbis viral RNA was infectious. Treatment of LAC virus with the nonionic detergent Triton X 100 and salt released three nucleocapsid structures, each containing one species of virion RNA. The nucleocapsids had sedimentation coefficients of 115S, 90S, and 65S. Negative contrast electron microscopy of the nucleocapsids indicated that they were convoluted, supercoiled, and apparently circular. They had a mean diameter of 10 to 12 nm and modal lengths of 200, 510, and 700 nm (some were even longer). By chemical and enzymatic analysis of purified viral RNA, one type of 5' nucleotide (pppAp) present in the proportion of one per RNA segment was identified. After periodate oxidation, each virion RNA species was labeled by reduction with [3H] sodium borohydride. Taken together, these results suggest that although the nucleocapsids appear as closed loops, the viral RNA has free 5' and 3' ends and is, therefore, not circular.",
author = "Obijeski, {J. F.} and Bishop, {D. H L} and Palmer, {E. L.} and Murphy, {F. A.}",
year = "1976",
language = "English (US)",
volume = "20",
pages = "664--675",
journal = "Journal of Virology",
issn = "0022-538X",
publisher = "American Society for Microbiology",
number = "3",

}

TY - JOUR

T1 - Segmented genome and nucleocapsid of La Crosse virus

AU - Obijeski, J. F.

AU - Bishop, D. H L

AU - Palmer, E. L.

AU - Murphy, F. A.

PY - 1976

Y1 - 1976

N2 - La Crosse (LAC) virions purified by velocity and equilibrium gradient centrifugation contained three single stranded RNA species. The three segments had sedimentation coefficients of 31S, 25S, and 12S by sodium dodecyl sulfate sucrose gradient centrifugation. By comparison with other viral and cellular RNA species, the LAC viral RNAs had molecular weights of 2.9x106, 1.8x106, and 0.4x106. Phenol sodium dodecyl sulfate extracted LAC virion RNA was not infectious for BHK 21 cell cultures under conditions in which Sindbis viral RNA was infectious. Treatment of LAC virus with the nonionic detergent Triton X 100 and salt released three nucleocapsid structures, each containing one species of virion RNA. The nucleocapsids had sedimentation coefficients of 115S, 90S, and 65S. Negative contrast electron microscopy of the nucleocapsids indicated that they were convoluted, supercoiled, and apparently circular. They had a mean diameter of 10 to 12 nm and modal lengths of 200, 510, and 700 nm (some were even longer). By chemical and enzymatic analysis of purified viral RNA, one type of 5' nucleotide (pppAp) present in the proportion of one per RNA segment was identified. After periodate oxidation, each virion RNA species was labeled by reduction with [3H] sodium borohydride. Taken together, these results suggest that although the nucleocapsids appear as closed loops, the viral RNA has free 5' and 3' ends and is, therefore, not circular.

AB - La Crosse (LAC) virions purified by velocity and equilibrium gradient centrifugation contained three single stranded RNA species. The three segments had sedimentation coefficients of 31S, 25S, and 12S by sodium dodecyl sulfate sucrose gradient centrifugation. By comparison with other viral and cellular RNA species, the LAC viral RNAs had molecular weights of 2.9x106, 1.8x106, and 0.4x106. Phenol sodium dodecyl sulfate extracted LAC virion RNA was not infectious for BHK 21 cell cultures under conditions in which Sindbis viral RNA was infectious. Treatment of LAC virus with the nonionic detergent Triton X 100 and salt released three nucleocapsid structures, each containing one species of virion RNA. The nucleocapsids had sedimentation coefficients of 115S, 90S, and 65S. Negative contrast electron microscopy of the nucleocapsids indicated that they were convoluted, supercoiled, and apparently circular. They had a mean diameter of 10 to 12 nm and modal lengths of 200, 510, and 700 nm (some were even longer). By chemical and enzymatic analysis of purified viral RNA, one type of 5' nucleotide (pppAp) present in the proportion of one per RNA segment was identified. After periodate oxidation, each virion RNA species was labeled by reduction with [3H] sodium borohydride. Taken together, these results suggest that although the nucleocapsids appear as closed loops, the viral RNA has free 5' and 3' ends and is, therefore, not circular.

UR - http://www.scopus.com/inward/record.url?scp=0017107005&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0017107005&partnerID=8YFLogxK

M3 - Article

C2 - 994302

AN - SCOPUS:0017107005

VL - 20

SP - 664

EP - 675

JO - Journal of Virology

JF - Journal of Virology

SN - 0022-538X

IS - 3

ER -