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Journal of Virology, December 2009, p. 12526-12534, Vol. 83, No. 23
0022-538X/09/$08.00+0     doi:10.1128/JVI.01219-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Packaging of Host mY RNAs by Murine Leukemia Virus May Occur Early in Y RNA Biogenesis{triangledown}

Eric L. Garcia,1 Adewunmi Onafuwa-Nuga,2 Soyeong Sim,3 Steven R. King,1 Sandra L. Wolin,3 and Alice Telesnitsky1*

Cellular and Molecular Biology Graduate Program and Department of Microbiology and Immunology, University of Michigan, Ann Arbor, Michigan,1 Department of Internal Medicine, University of Michigan, Ann Arbor, Michigan,2 Departments of Cell Biology and Molecular Biophysics and Biochemistry, Yale University School of Medicine, New Haven, Connecticut3

Received 12 June 2009/ Accepted 15 September 2009

Moloney murine leukemia virus (MLV) selectively encapsidates host mY1 and mY3 RNAs. These noncoding RNA polymerase III transcripts are normally complexed with the Ro60 and La proteins, which are autoantigens associated with rheumatic disease that function in RNA biogenesis and quality control. Here, MLV replication and mY RNA packaging were analyzed using Ro60 knockout embryonic fibroblasts, which contain only ~3% as much mY RNA as wild-type cells. Virus spread at the same rate in wild-type and Ro knockout cells. Surprisingly, MLV virions shed by Ro60 knockout cells continued to package high levels of mY1 and mY3 (about two copies of each) like those from wild-type cells, even though mY RNAs were barely detectable within producer cells. As a result, for MLV produced in Ro60 knockout cells, encapsidation selectivity from among all cell RNAs was even higher for mY RNAs than for the viral genome. Whereas mY RNAs are largely cytoplasmic in wild-type cells, fractionation of knockout cells revealed that the residual mY RNAs were relatively abundant in nuclei, likely reflecting the fact that most mY RNAs were degraded shortly after transcription in the absence of Ro60. Together, these data suggest that these small, labile host RNAs may be recruited at a very early stage of their biogenesis and may indicate an intersection of retroviral assembly and RNA quality control pathways.

* Corresponding author. Mailing address: 1150 W. Medical Center Dr., Room 5641, Ann Arbor, MI 48109-5620. Phone: (734) 936-6466. Fax: (734) 764-3562. E-mail: ateles{at}umich.edu

{triangledown} Published ahead of print on 23 September 2009.

Journal of Virology, December 2009, p. 12526-12534, Vol. 83, No. 23
0022-538X/09/$08.00+0     doi:10.1128/JVI.01219-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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