Evelyn Kilareski Research Instructor, Microbiology and Immunology Ph.D., Pennsylvania State University College of Medicine, 2003 245 N. 15th Street Philadelphia, PA  19102 Tel: 215-762-4139 Fax: 215-762-7784 Email:  evelyn.kilareski@drexelmed.edu

Keywords:

Human Immunodeficiency virus, HIV-1, transcription, chromatin, LTR

HIV-1 has an exceptionally high frequency of genetic variation within infected individuals due to the high error rate of the HIV-1 reverse transcriptase enzyme. These variations occur in both the coding region of the HIV-1 genome and in the promoter, or long terminal repeat (LTR). Work in the Wigdahl laboratory has identified several genetic variations within the LTR of HIV-1 that correlate with HIV-associated disease progression, and specifically HIV-associated dementia (HIVD).  

The frequency of a C to T change at position 3 of C/EBP binding site I increases in the peripheral blood of patients as HIV-1-associated disease progresses. Likewise, a C to T change at position 5 of Sp binding site III in the LTR also increases in frequency as disease progresses. Interestingly, these variants are only found in the brain tissue of patients who suffer from HIVD, and not in the brain tissue of AIDS patients who do not have HIVD. These genetic variations have been shown to alter transcription factor recruitment in vitro, but the effect of these in vivo has not been determined.

My work in the laboratory of Dr. Brian Wigdahl is to continue to investigate the correlation between HIV-1 sequence variation and AIDS-related neurological dysfunction, and to determine the effect of LTR variation on factor recruitment in vivo. One aspect of this work is to study the effect of LTR variations on viral replication. To do so, the LTR variations are incorporated into infectious molecular clones corresponding to CXCR4-utilizing, CCR5-utilizing, or dual tropic HIV-1. The resulting viral particles are then used in infection studies in both cell lines and primary peripheral blood mononuclear cells. A second, related aspect of this work is to determine the molecular mechanism behind differences seen in viral replication caused by genetic changes in the LTR. This is done using chromatin immunoprecipitation and electrophoretic mobility shift assays.

Selected Publications:

1. Springhetti, E.M, N.E., Istomina, C. Woodcock, J. Whisstock, S.A. Grigoryev. 2003. Bridging and folding of nucleosome arrays by MENT: the role of the M-loop and RCL. The Journal of Biological Chemistry. 278(44): 43384-43393.
   
   
2. Istomina, N.E.*, S.S. Shushanov*, E.M. Springhetti*, V.L. Karpov, I.A. Krasheninnikov, K. Stevens, K.S. Zaret, P.B. Singh, and S.A. Grigoryev. 2003. Insulation of the chicken ƒÀ-globin chromosomal domain from a chromatin-condensing protein, MENT. Molecular and Cellular Biology. 23(18):6455-6468. *Authors contributed equally.


3. Alexaki, A., Quiterio, S., Liu, Y., Irish, B., Kilareski, E., Nonnemacher, M., and B. Wigdahl.  PMA-induced differentiation of a bone marrow progenitor cell line activates HIV-1 LTR-driven transcription.  DNA and Cell Biology, 26(6): 387-394, 2007.


4. Alexaki, A., Banerjee, A., Kilareski, E., Nonnemacher, M. R., and B. Wigdahl.  IL-1β production by differentiating TF-1 bone marrow progenitor cells.  Proceedings of the 8th International Congress of Neuroimmunology, 353-359, 2007.