This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Li, L.
Right arrow Articles by Bushman, F. D.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Li, L.
Right arrow Articles by Bushman, F. D.

 Previous Article  |  Next Article 

J Virol, March 1998, p. 2125-2131, Vol. 72, No. 3
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Modulation of Activity of Moloney Murine Leukemia Virus Preintegration Complexes by Host Factors In Vitro

Ling Li,1 Chris M. Farnet,2,dagger W. French Anderson,1 and Frederic D. Bushman2,*

Infectious Disease Laboratory, Salk Institute for Biological Studies, La Jolla, California 92037,2 and Gene Therapy Laboratories, Norris Cancer Center, University of Southern California School of Medicine, Los Angeles, California 900331

Received 15 September 1997/Accepted 25 November 1997

We have explored the requirements for host proteins in the integration of Moloney murine leukemia virus (MoMuLV) cDNA in vitro. Following infection, it is possible to lyse cells and obtain preintegration complexes (PICs) capable of integrating the MoMuLV cDNA into an added target DNA in vitro (intermolecular integration). PICs can be stripped of required proteins by gel filtration in high-salt buffers (600 mM KCl), allowing the nature of the removed factors to be investigated by in vitro reconstitution. In a previous study of human immunodeficiency virus type 1 (HIV-1) PICs, the host protein HMG I(Y) was found to be able to restore activity to salt-stripped PICs. In contrast, salt stripping and reconstitution of MoMuLV PICs led to the proposal that a host factor is important for a different activity, blocking integration into the cDNA itself (autointegration). In this report, we investigated reconstitution of salt-stripped MoMuLV PICs and found that addition of cellular extract from uninfected NIH 3T3 cells could block autointegration and also restore intermolecular integration. Isolation of the intermolecular integration-complementing activity yielded HMG I(Y), as in the HIV-1 case. However, HMG I(Y) could not block autointegration, implicating a different host factor in this process. Additionally, when MoMuLV PICs were partially purified but not salt stripped, the intermolecular integration activity was reduced but could be stimulated by the addition of any of several purified DNA binding proteins. In summary, three activities were detected: (i) the intermolecular integration cofactor HMG I(Y), (ii) an autointegration barrier protein, and (iii) stimulatory DNA binding proteins.

* Corresponding author. Mailing address: Infectious Disease Laboratory, Salk Institute for Biological Studies, 10010 N. Torrey Pines Rd., La Jolla, CA 92037. Phone: (619) 453-4100. Fax: (619) 554-0341. E-mail: rick_bushman{at}qm.salk.edu.

dagger Present address: Bio-Mega Research Division of Boehringer Ingelheim, Canada, Ltd., Laval, Quebec H7S 2G5, Canada.




This article has been cited by other articles:

  • Puglia, J., Wang, T., Smith-Snyder, C., Cote, M., Scher, M., Pelletier, J. N., John, S., Jonsson, C. B., Roth, M. J. (2006). Revealing Domain Structure through Linker-Scanning Analysis of the Murine Leukemia Virus (MuLV) RNase H and MuLV and Human Immunodeficiency Virus Type 1 Integrase Proteins. J. Virol. 80: 9497-9510 [Abstract] [Full Text]  
  • Jarrosson-Wuilleme, L., Goujon, C., Bernaud, J., Rigal, D., Darlix, J.-L., Cimarelli, A. (2006). Transduction of Nondividing Human Macrophages with Gammaretrovirus-Derived Vectors. J. Virol. 80: 1152-1159 [Abstract] [Full Text]  
  • Bruce, J. W., Bradley, K. A., Ahlquist, P., Young, J. A. T. (2005). Isolation of Cell Lines That Show Novel, Murine Leukemia Virus-Specific Blocks to Early Steps of Retroviral Replication. J. Virol. 79: 12969-12978 [Abstract] [Full Text]  
  • Mansharamani, M., Graham, D. R. M., Monie, D., Lee, K. K., Hildreth, J. E. K., Siliciano, R. F., Wilson, K. L. (2003). Barrier-to-Autointegration Factor BAF Binds p55 Gag and Matrix and Is a Host Component of Human Immunodeficiency Virus Type 1 Virions. J. Virol. 77: 13084-13092 [Abstract] [Full Text]  
  • Lin, C.-W., Engelman, A. (2003). The Barrier-to-Autointegration Factor Is a Component of Functional Human Immunodeficiency Virus Type 1 Preintegration Complexes. J. Virol. 77: 5030-5036 [Abstract] [Full Text]  
  • Gao, K., Gorelick, R. J., Johnson, D. G., Bushman, F. (2002). Cofactors for Human Immunodeficiency Virus Type 1 cDNA Integration In Vitro. J. Virol. 77: 1598-1603 [Abstract] [Full Text]  
  • Suzuki, Y., Craigie, R. (2002). Regulatory Mechanisms by Which Barrier-to-Autointegration Factor Blocks Autointegration and Stimulates Intermolecular Integration of Moloney Murine Leukemia Virus Preintegration Complexes. J. Virol. 76: 12376-12380 [Abstract] [Full Text]  
  • Yuan, B., Fassati, A., Yueh, A., Goff, S. P. (2002). Characterization of Moloney Murine Leukemia Virus p12 Mutants Blocked during Early Events of Infection. J. Virol. 76: 10801-10810 [Abstract] [Full Text]  
  • Serhan, F., Jourdan, N., Saleun, S., Moullier, P., Duisit, G. (2002). Characterization of Producer Cell-Dependent Restriction of Murine Leukemia Virus Replication. J. Virol. 76: 6609-6617 [Abstract] [Full Text]  
  • Sinha, S., Pursley, M. H., Grandgenett, D. P. (2002). Efficient Concerted Integration by Recombinant Human Immunodeficiency Virus Type 1 Integrase without Cellular or Viral Cofactors. J. Virol. 76: 3105-3113 [Abstract] [Full Text]  
  • Dirks, C., Miller, A. D. (2001). Many Nonmammalian Cells Exhibit Postentry Blocks to Transduction by Gammaretroviruses Pseudotyped with Various Viral Envelopes, Including Vesicular Stomatitis Virus G Glycoprotein. J. Virol. 75: 6375-6383 [Abstract] [Full Text]  
  • Vora, A., Grandgenett, D. P. (2001). DNase Protection Analysis of Retrovirus Integrase at the Viral DNA Ends for Full-Site Integration In Vitro. J. Virol. 75: 3556-3567 [Abstract] [Full Text]  
  • Li, L., Yoder, K., Hansen, M. S. T., Olvera, J., Miller, M. D., Bushman, F. D. (2000). Retroviral cDNA Integration: Stimulation by HMG I Family Proteins. J. Virol. 74: 10965-10974 [Abstract] [Full Text]  
  • Chiu, R., Grandgenett, D. P. (2000). Avian Retrovirus DNA Internal Attachment Site Requirements for Full-Site Integration In Vitro. J. Virol. 74: 8292-8298 [Abstract] [Full Text]  
  • Szurek, P. F., Brooks, B. R. (2000). Synthesis of Virus-Specific High-Mobility DNA after Temperature Upshift of SC-1 Cells Chronically Infected with Moloney Murine Leukemia Virus Mutant ts1. J. Virol. 74: 7055-7063 [Abstract] [Full Text]  
  • Carteau, S., Gorelick, R. J., Bushman, F. D. (1999). Coupled Integration of Human Immunodeficiency Virus Type 1 cDNA Ends by Purified Integrase In Vitro: Stimulation by the Viral Nucleocapsid Protein. J. Virol. 73: 6670-6679 [Abstract] [Full Text]  
  • Leblanc, P., Dastugue, B., Vaury, C. (1999). The Integration Machinery of ZAM, a Retroelement from Drosophila melanogaster, Acts as a Sequence-Specific Endonuclease. J. Virol. 73: 7061-7064 [Abstract] [Full Text]  
  • Bustin, M. (1999). Regulation of DNA-Dependent Activities by the Functional Motifs of the High-Mobility-Group Chromosomal Proteins. Mol. Cell. Biol. 19: 5237-5246 [Full Text]  
  • Zhao, Y., Zhu, L., Lee, S., Li, L., Chang, E., Soong, N.-W., Douer, D., Anderson, W. F. (1999). Identification of the block in targeted retroviral-mediated gene transfer. Proc. Natl. Acad. Sci. USA 96: 4005-4010 [Abstract] [Full Text]  
  • Chen, H., Engelman, A. (1998). The barrier-to-autointegration protein is a host factor for HIV type 1 integration. Proc. Natl. Acad. Sci. USA 95: 15270-15274 [Abstract] [Full Text]  
  • Harris, D., Engelman, A. (2000). Both the Structure and DNA Binding Function of the Barrier-to-Autointegration Factor Contribute to Reconstitution of HIV Type 1 Integration in Vitro. J. Biol. Chem. 275: 39671-39677 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»