Faculty Information
Michael A. Belshan, Ph.D.
Appointment & Titles |  Education |  Research and Professional Experience
Honors & Awards  |  Research Interests |  Selected Publications  |  Links |  Contact

Appointment(s) and Titles:
Assistant Professor, Medical Microbiology and Immunology, Creighton University
Faculty, Nebraska Center for Virology, University of Nebraska, Lincoln, NE


B.S., 1993, Zoology, Iowa State University, Ames IA

Ph.D., 1999, Molecular, Cellular, and Developmental Biology, and Microbiology, Iowa State University, Ames IA

Research and Professional Experience:

Postdoctoral Fellow, 2000-2005, Molecular Oncology, Washington University School of Medicine, St. Louis MO

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Honors and Awards:

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Research Interests:

HIV Molecular Virology

 Human Immunodeficiency Virus (HIV) is the causative agent of acquired immunodeficiency syndrome (AIDS). HIV infection is incurable, but treatment with a combination of anti-retroviral drugs can reduce virus replication to undetectable levels and forestall disease progression.

 However drug resistance to the existing pool of anti-retroviral therapies continues to rise. Continued success in the repression of HIV replication in infected individuals will require the development of new inhibitors that are effective against drug-resistant strains of virus. Drugs that target novel areas of virus replication have the greatest probability to be effective against such viruses.

 Our laboratory seeks to comprehend poorly understood aspects of HIV replication to identify new targets for anti-viral therapies. Several areas of HIV replication are potential targets. These include virus uncoating, preintegration complex (PIC) assembly and transport, viral transcriptional regulation, and virus assembly and release.

 Currently we are focused on identifying novel cellular components of PICs as a first step to develop a new class of inhibitors. PICs are large viral DNA (vDNA) complexes formed during early HIV-1 infection of cells. They facilitate the transport of viral DNA into the nucleus of cells. Due to difficulties in producing and purifying PICs, their composition, assembly, and transport remain poorly defined. The viral proteins associated with the PIC do not account for the size of the complexes, suggesting they contain additional cellular components. Our long-term goal is to aid in the development of new HIV-1 inhibitors by determining how PICs assemble and mediate nuclear transport of vDNA in dividing and non-dividing cells.

 Disparities exist in the literature concerning methods to both produce and purify preintegration complexes (PICs). We have analyzed several methods of PIC production and purification and developed a method that consistently produces PICs from several cell types using velocity gradient centrifugation. We have characterized the biochemical properties of these complexes and identified the location of PICs in gradients. Our protocol produces PICs that: 1) are biochemically similar to in vivo PICs; 2) are of sufficient quantity to analyze by mass spectrometry; and 3) are active for nuclear transport and integration in vitro.

 In parallel studies in collaboration with Dr. Alan Engelman at the Dana Farber Cancer Institute we have developed a method to biotinylate HIV PIC proteins in vivo. Biotinylated proteins can be affinity purified with streptavidin to capture protein complexes and identify novel cellular proteins that interact with HIV.

 Our mass spectrometry analysis is done in collaboration with Dr. Pawel Ciborowski, the Director of the Mass Spectrometry Core Facility at the University of Nebraska Medical Center, and Dr. Ron Cerny, the Director of the University of Nebraska at Lincoln Mass Spectrometry Facility. Both are members of the Nebraska Center for Virology.

 Our MS analyses consistently identify viral proteins in every infected sample. We have also detected a number of cellular proteins previously demonstrated to interact with HIV-1 PICs and/or modulate HIV-1 replication. Several potential candidate proteins have been identified using both the targeted and affinity purification approaches. Four candidate proteins have been detected in virus particles. We are now working to characterize the role of these proteins in HIV replication and we continue to discover novel cellular components of PICs that play a critical role in retroviral replication.

 Our laboratory is affiliated with the Nebraska Center for Virology a Center of Biomedical Research Excellence formed in the fall of 2000 under the National Institutes of Health Institutional Development Award program. The Center links the virology programs of Creighton University, the University of Nebraska at Lincoln, and the University of Nebraska Medical Center and conducts innovative research that addresses fundamental questions about the pathogenesis and replication of diverse viral agents that effect human, animal, and plant health.

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Selected Publicatons:

Belshan, M., M.E. Harris, A.E. Shoemaker, T.J. Hope, and S. Carpenter. 1998. Biological characterization of rev variation in equine infectious anemia virus. J. Virol. 72:4421-4426.

 Belshan, M., G. Park, P. Bilodeau, C.M. Stoltzfus, and S. Carpenter. 2000. Binding of equine infectious anemia virus Rev to an exon splicing enhancer mediates alternative splicing and nuclear export of viral mRNAs. Mol. Cell Biol. 20:3550-3557.

 Belshan, M., P. Baccam, J.L. Oaks, S. Murphy, J.L. Cornette, and S. Carpenter. 2001. Genetic and biological variation in equine infectious anemia virus Rev at different stages of clinical disease:  Implications for virus persistence. Virology. 279:185-200.

 Belshan, M. and L. Ratner. 2003. Identification of the nuclear localization signal of human immunodeficiency virus type 2 Vpx. Virology. 311:7-15.

 Baccam, P., R.J. Thompson, Y. Li, W.O. Sparks, M. Belshan, K.S. Dorman, Y. Wannemuehler, J.L. Oaks, J.L. Cornette, and S. Carpenter. 2003. Subpopulations of equine infectious anemia virus Rev co-exist in vivo and differ in phenotype. J. Virol. 77:12122-31.

 Belshan, M., L. Mahnke., and L. Ratner. 2005. Conserved amino acids are critical components of the nuclear localization signal of human immunodeficiency virus type 2 Vpx. Virology. 346:118-26.

 Mahnke, L.A., M. Belshan, and L. Ratner. 2006. Analysis of HIV-2 Vpx by modeling and insertional mutagenesis. Virology. 348:165-74.

 Lee, J.-H., S.C. Murphy, M. Belshan, W.O. Sparks, Y. Wannemuehler, S. Liu, D. Dobbs, T.J. Hope, D. Dobbs, and S. Carpenter. 2006. Characterization of functional domains of equine infectious anemia virus Rev suggests a bipartite RNA-binding domain. J. Virol. 80:3844-52.

 Cheng, X., M. Belshan, and L. Ratner. 2007. Hsp40 facilitates HIV-2 Vpx-mediated preintegration complex nuclear import. J. Virol. 82:1229-37.



Nebraska Center for Virology

American Society for Virology

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