Role of cellular FKBP52 protein in intracellular trafficking of recombinant adeno-associated virus 2 vectors

Weihong Zhao, Li Zhong, Jianqing Wu, Linyuan Chen, Keyun Qing, Kirsten A. Weigel-Kelley, Steven H. Larsen, Weinian Shou, Kenneth H. Warrington, Arun Srivastava

Research output: Contribution to journalArticle

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Abstract

We have reported that tyrosine-phosphorylated forms of a cellular protein, FKBP52, inhibit the second-strand DNA synthesis of adeno-associated virus 2 (AAV), leading to inefficient transgene expression from recombinant AAV vectors. To further explore the role of FKBP52 in AAV-mediated transduction, we established murine embryo fibroblasts (MEFs) cultures from FKBP52 wild-type (WT), heterozygous (HE), and knockout (KO) mice. Conventional AAV vectors failed to transduce WT MEFs efficiently, and the transduction efficiency was not significantly increased in HE or KO MEFs. AAV vectors failed to traffic efficiently to the nucleus in these cells. Treatment with hydroxyurea (HU) increased the transduction efficiency of conventional AAV vectors by ∼25-fold in WT MEFs, but only by ∼4-fold in KO MEFs. The use of self-complementary AAV (scAAV) vectors, which bypass the requirement of viral second-strand DNA synthesis, revealed that HU treatment increased the transduction efficiency ∼23-fold in WT MEFs, but only ∼4-fold in KO MEFs, indicating that the lack of HU treatment-mediated increase in KO MEFs was not due to failure of AAV to undergo viral second-strand DNA synthesis. Following HU treatment, ∼59% of AAV genomes were present in the nuclear fraction from WT MEFs, but only ∼28% in KO MEFs, indicating that the pathway by which HU treatment mediates nuclear transport of AAV was impaired in KO MEFs. When KO MEFs were stably transfected with an FKBP52 expression plasmid, HU treatment-mediated increase in the transduction efficiency was restored in these cells, which correlated directly with improved intracellular trafficking. Intact AAV particles were also shown to interact with FKBP52 as well as with dynein, a known cellular protein involved in AAV trafficking. These studies suggest that FKBP52, being a cellular chaperone protein, facilitates intracellular trafficking of AAV, which has implications in the optimal use of recombinant AAV vectors in human gene therapy.

Original languageEnglish
Pages (from-to)283-293
Number of pages11
JournalVirology
Volume353
Issue number2
DOIs
StatePublished - Sep 30 2006

Fingerprint

Dependovirus
Embryonic Structures
Fibroblasts
Hydroxyurea
Proteins
tacrolimus binding protein 4
DNA
Therapeutics
Dyneins
Cell Nucleus Active Transport
Cell Nucleus
Transgenes
Knockout Mice
Genetic Therapy
Virion
Tyrosine

Keywords

  • Adeno-associated virus type 2
  • FK506-binding protein
  • Gene expression
  • Gene transfer
  • Intracellular trafficking
  • Murine embryo fibroblasts
  • Viral vectors

ASJC Scopus subject areas

  • Virology
  • Infectious Diseases

Cite this

Zhao, W., Zhong, L., Wu, J., Chen, L., Qing, K., Weigel-Kelley, K. A., ... Srivastava, A. (2006). Role of cellular FKBP52 protein in intracellular trafficking of recombinant adeno-associated virus 2 vectors. Virology, 353(2), 283-293. https://doi.org/10.1016/j.virol.2006.04.042

Role of cellular FKBP52 protein in intracellular trafficking of recombinant adeno-associated virus 2 vectors. / Zhao, Weihong; Zhong, Li; Wu, Jianqing; Chen, Linyuan; Qing, Keyun; Weigel-Kelley, Kirsten A.; Larsen, Steven H.; Shou, Weinian; Warrington, Kenneth H.; Srivastava, Arun.

In: Virology, Vol. 353, No. 2, 30.09.2006, p. 283-293.

Research output: Contribution to journalArticle

Zhao, W, Zhong, L, Wu, J, Chen, L, Qing, K, Weigel-Kelley, KA, Larsen, SH, Shou, W, Warrington, KH & Srivastava, A 2006, 'Role of cellular FKBP52 protein in intracellular trafficking of recombinant adeno-associated virus 2 vectors', Virology, vol. 353, no. 2, pp. 283-293. https://doi.org/10.1016/j.virol.2006.04.042
Zhao, Weihong ; Zhong, Li ; Wu, Jianqing ; Chen, Linyuan ; Qing, Keyun ; Weigel-Kelley, Kirsten A. ; Larsen, Steven H. ; Shou, Weinian ; Warrington, Kenneth H. ; Srivastava, Arun. / Role of cellular FKBP52 protein in intracellular trafficking of recombinant adeno-associated virus 2 vectors. In: Virology. 2006 ; Vol. 353, No. 2. pp. 283-293.
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