Knockdown of the cellular protein lrpprc attenuates HIV-1 infection

Cameron J. Schweitzer, John M. Matthews, Christian J. Madson, Meghan R. Donnellan, Ronald L. Cerny, Michael A. Belshan

Research output: Contribution to journalArticle

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Abstract

HIV-1 exploits numerous host cellular pathways for productive infection. To identify novel factors involved in HIV-1 replication, HIV-1 integrase and matrix protein complexes were captured at 4 hours post infection for proteomic analysis using an affinity purification system. Leucine-rich PPR-motif containing (LRPPRC) protein, a cellular protein involved in mitochondrial function, cell metabolism, and cell-cycle progression was identified as one of the candidate HIV-1 factors. Co-immunoprecipitation RT-PCR experiments confirmed that LRPPRC associated with HIV-1 nucleic acids during the early steps of virus infection. To establish if LRPPRC was critical for HIV-1 infection, three independent LRPPRC knockdown cell lines were constructed (2.7, 3.6, and 4.1). Subcellular fractionation of these cell lines revealed differential knockdown of LRPPRC in subcellular compartments. LRPPRC was knocked down in the insoluble/cytoskeletal fractions of all three cell lines, but the 3.6 and 4.1 cells also showed a reduction in nuclear LRPPRC. Additionally, several cellular factors were downregulated and/or disrupted by loss of LRPPRC. HIV-1 infection was reduced in all three cell lines, but virus production and RNA encapsidation were unaffected, suggesting that LRPPRC was critical for the afferent stage of virus replication. Two of the three cell lines (3.6, 4.1) were refractory for murine leukemia virus infection, a virus dependent on cellular proliferation for productive infection. Consistent with this, these two cell lines exhibited reduced cellular growth with no loss of cellular viability or change in cell cycle phenotype. The early steps of virus infection were also differentially affected among the cell lines. A reduced level of preintegration complex formation was observed in all three cell lines, but viral DNA nuclear import was reduced only in the 3.6 and 4.1 cells. Combined, these data identify LRPPRC as a HIV-1 factor that is involved in HIV-1 replication through more than one mechanism.

Original languageEnglish
Article numbere40537
JournalPLoS One
Volume7
Issue number7
DOIs
StatePublished - Jul 12 2012

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Human immunodeficiency virus 1
Leucine
leucine
HIV Infections
HIV-1
Cells
Viruses
cell lines
Cell Line
infection
Proteins
proteins
Virus Diseases
viruses
cell cycle
Cell Cycle
Infection
Murine leukemia virus
Murine Leukemia Viruses
Amino Acid Motifs

All Science Journal Classification (ASJC) codes

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Schweitzer, C. J., Matthews, J. M., Madson, C. J., Donnellan, M. R., Cerny, R. L., & Belshan, M. A. (2012). Knockdown of the cellular protein lrpprc attenuates HIV-1 infection. PLoS One, 7(7), [e40537]. https://doi.org/10.1371/journal.pone.0040537

Knockdown of the cellular protein lrpprc attenuates HIV-1 infection. / Schweitzer, Cameron J.; Matthews, John M.; Madson, Christian J.; Donnellan, Meghan R.; Cerny, Ronald L.; Belshan, Michael A.

In: PLoS One, Vol. 7, No. 7, e40537, 12.07.2012.

Research output: Contribution to journalArticle

Schweitzer, CJ, Matthews, JM, Madson, CJ, Donnellan, MR, Cerny, RL & Belshan, MA 2012, 'Knockdown of the cellular protein lrpprc attenuates HIV-1 infection', PLoS One, vol. 7, no. 7, e40537. https://doi.org/10.1371/journal.pone.0040537
Schweitzer CJ, Matthews JM, Madson CJ, Donnellan MR, Cerny RL, Belshan MA. Knockdown of the cellular protein lrpprc attenuates HIV-1 infection. PLoS One. 2012 Jul 12;7(7). e40537. https://doi.org/10.1371/journal.pone.0040537
Schweitzer, Cameron J. ; Matthews, John M. ; Madson, Christian J. ; Donnellan, Meghan R. ; Cerny, Ronald L. ; Belshan, Michael A. / Knockdown of the cellular protein lrpprc attenuates HIV-1 infection. In: PLoS One. 2012 ; Vol. 7, No. 7.
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