Abstract:
:Adeno-associated virus (AAV) is gaining momentum as a gene therapy vector for human applications. However, there remain impediments to the development of this virus as a vector. One of these is the incomplete understanding of the biology of the virus, including nuclear targeting of the incoming virion during initial infection, as well as assembly of progeny virions from structural components in the nucleus. Toward this end, we have identified four basic regions (BR) on the AAV2 capsid that represent possible nuclear localization sequence (NLS) motifs. Mutagenesis of BR1 ((120)QAKKRVL(126)) and BR2 ((140)PGKKRPV(146)) had minor effects on viral infectivity ( approximately 4- and approximately 10-fold, respectively), whereas BR3 ((166)PARKRLN(172)) and BR4 ((307)RPKRLN(312)) were found to be essential for infectivity and virion assembly, respectively. Mutagenesis of BR3, which is located in Vp1 and Vp2 capsid proteins, does not interfere with viral production or trafficking of intact AAV capsids to the nuclear periphery but does inhibit transfer of encapsidated DNA into the nucleus. Substitution of the canine parvovirus NLS rescued the BR3 mutant to wild-type (wt) levels, supporting the role of an AAV NLS motif. In addition, rAAV2 containing a mutant form of BR3 in Vp1 and a wt BR3 in Vp2 was found to be infectious, suggesting that the function of BR3 is redundant between Vp1 and Vp2 and that Vp2 may play a role in infectivity. Mutagenesis of BR4 was found to inhibit virion assembly in the nucleus of transfected cells. This affect was not completely due to the inefficient nuclear import of capsid subunits based on Western blot analysis. In fact, aberrant capsid foci were observed in the cytoplasm of transfected cells, compared to the wild type, suggesting a defect in early viral assembly or trafficking. Using three-dimensional structural analysis, the lysine- and arginine-to-asparagine change disrupts hydrogen bonding between these basic residues and adjacent beta strand glutamine residues that may prevent assembly of intact virions. Taken together, these data support that the BR4 domain is essential for virion assembly. Each BR was also found to be conserved in serotypes 1 to 11, suggesting that these regions are significant and function similarly in each serotype. This study establishes the importance of two BR motifs on the AAV2 capsid that are essential for infectivity and virion assembly.
journal_name
J Viroljournal_title
Journal of virologyauthors
Grieger JC,Snowdy S,Samulski RJdoi
10.1128/JVI.02723-05subject
Has Abstractpub_date
2006-06-01 00:00:00pages
5199-210issue
11eissn
0022-538Xissn
1098-5514pii
80/11/5199journal_volume
80pub_type
杂志文章abstract::To examine the prerequisites for cleavage activation of the hemagglutinin of human influenza viruses, a cDNA clone obtained from strain A/Port Chalmers/1/73 (serotype H3) was subjected to site-directed mutagenesis and expressed in CV-1 cells by using a simian virus 40 vector. The number of basic residues at the cleava...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.65.7.3530-3537.1991
更新日期:1991-07-01 00:00:00
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journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.63.5.2411-2414.1989
更新日期:1989-05-01 00:00:00
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pub_type: 杂志文章
doi:10.1128/JVI.72.10.8181-8190.1998
更新日期:1998-10-01 00:00:00
abstract::Five minority populations of aberrant, varicella-zoster virus (VZV)-derived genomes were identified among the encapsidated DNAs obtained from the nuclear and cytoplasmic fractions of an in vitro infection initiated with a lyophilized sample of the BIKEN VZV vaccine (strain Oka). These were (i) VZV genomes, present wit...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.56.1.92-101.1985
更新日期:1985-10-01 00:00:00
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journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.28.3.997-1001.1978
更新日期:1978-12-01 00:00:00
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journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/jvi.75.17.8147-8157.2001
更新日期:2001-09-01 00:00:00
abstract::Our previous study has shown that recombinant adeno-associated virus (rAAV) vector integrates preferentially in genes, near transcription start sites and CpG islands in mouse liver (H. Nakai, X. Wu, S. Fuess, T. A. Storm, D. Munroe, E. Montini, S. M. Burgess, M. Grompe, and M. A. Kay, J. Virol. 79:3606-3614, 2005). Ho...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.00963-07
更新日期:2007-10-01 00:00:00
abstract::Understanding how viral components collaborate to convert the human immunodeficiency virus type 1 genome from single-stranded RNA into double-stranded DNA is critical to the understanding of viral replication. Not only must the correct reactions be carried out, but unwanted side reactions must be avoided. After minus-...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/jvi.74.19.8785-8792.2000
更新日期:2000-10-01 00:00:00
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journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.06113-11
更新日期:2012-02-01 00:00:00
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journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.02891-12
更新日期:2013-03-01 00:00:00
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journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.64.2.886-889.1990
更新日期:1990-02-01 00:00:00
abstract::Epstein-Barr virus (EBV) in vivo is known to establish persistent infection in resting, circulating memory B cells and to productively replicate in plasma cells. Until now, the molecular mechanism of how EBV switches from latency to lytic replication in vivo was not known. Here, we report that the plasma cell differen...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.01055-07
更新日期:2007-12-01 00:00:00
abstract:UNLABELLED:SSV-type integrases, encoded by fuselloviruses which infect the hyperthermophilic archaea of the Sulfolobales, are archaeal members of the tyrosine recombinase family. These integrases catalyze viral integration into and excision from a specific site on the host genome. In the present study, we have establis...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.01637-15
更新日期:2015-11-01 00:00:00
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journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.16.3.479-485.1975
更新日期:1975-09-01 00:00:00
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journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.7.1.176-187.1971
更新日期:1971-01-01 00:00:00
abstract::We cloned and analyzed the integrated human papillomavirus type 16 (HPV-16) genomes that are present in the human cervical carcinoma cell lines SiHa and CaSki. The single HPV-16 genome in the SiHa line was cloned as a 10-kilobase (kb) HindIII fragment. Integration of the HPV-16 genome occurred at bases 3132 and 3384 w...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.61.4.962-971.1987
更新日期:1987-04-01 00:00:00
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journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.69.3.1480-1484.1995
更新日期:1995-03-01 00:00:00
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journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.60.2.423-430.1986
更新日期:1986-11-01 00:00:00
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journal_title:Journal of virology
pub_type: 杂志文章
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更新日期:2011-12-01 00:00:00
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journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.79.11.7005-7013.2005
更新日期:2005-06-01 00:00:00
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journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.72.12.9470-9478.1998
更新日期:1998-12-01 00:00:00
abstract::Proliferation responses of naïve CD4(+) T cells to T-cell receptor and interleukin-7 (IL-7) stimulation were evaluated by using cells from human immunodeficiency virus-positive (HIV(+)) donors. IL-7 enhanced responses to T-cell receptor stimulation, and the magnitude of this enhancement was similar in cells from healt...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.00476-07
更新日期:2007-11-01 00:00:00
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journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.71.12.9482-9489.1997
更新日期:1997-12-01 00:00:00
abstract::The nef genes of the human immunodeficiency viruses type 1 and 2 (HIV-1 and HIV-2) and the related simian immunodeficiency viruses (SIVs) encode a protein (Nef) whose role in virus replication and cytopathicity remains uncertain. As an attempt to elucidate the function of nef, we characterized the nucleotide and corre...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.67.8.4639-4650.1993
更新日期:1993-08-01 00:00:00
abstract::Polyoma virus particles were purified by equilibrium centrifugation in CsCl. Particles from three regions of the density gradient were examined for infectivity, for their ability to induce expanded pools of deoxyribonucleic acid (DNA) precursors, and for their ability to stimulate the synthesis of DNA. The most infect...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.6.1.28-32.1970
更新日期:1970-07-01 00:00:00
abstract::Here, we investigated the pre-steady-state deoxynucleoside triphosphate (dNTP) incorporation kinetics of primate foamy virus (PFV) reverse transcriptase (RT) in comparison with those of HIV-1 and MuLV RTs. PFV RT displayed a drastic reduction in primer extension at low dNTP concentrations where HIV-1 RT remains highly...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.00088-08
更新日期:2008-08-01 00:00:00
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journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.71.3.2252-2263.1997
更新日期:1997-03-01 00:00:00
abstract::In clinical trials, HIV-1 broadly neutralizing antibodies (bnAbs) effectively lower plasma viremia and delay virus reemergence. The presence of less neutralization-susceptible strains prior to treatment decreases the efficacy of these antibody-based treatments, but neutralization sensitivity often cannot be predicted ...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.01604-19
更新日期:2020-01-06 00:00:00
abstract::The Cp promoter of Epstein-Barr virus (EBV) directs most transcription of the EBNA genes in lymphoblastoid cell lines. The functions of two control regions in the Cp promoter have been studied by construction of recombinant EBV strains containing specific mutations in these elements. Mutation of the RBP-Jk (CBF1) bind...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.70.3.1695-1705.1996
更新日期:1996-03-01 00:00:00
abstract:UNLABELLED:An altered intestinal microbiome during chronic human immunodeficiency virus (HIV) infection is associated with mucosal dysfunction, inflammation, and disease progression. We performed a preclinical evaluation of the safety and efficacy of fecal microbiota transplantation (FMT) as a potential therapeutic in ...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.00099-16
更新日期:2016-04-29 00:00:00