Overview

I grew up in Montclair NJ, received my BS from Douglass College/Rutgers University, and completed a senior thesis under
the wonderful mentorship of Dr. Evelyn Witkin. After a year working as a Research Technician with Dr. Richard Burgess
at the U. of Wisconsin, I joined Dr. Lucy Shapiro's laboratory at Einstein College of Medicine for my PhD work in
Molecular Biology and Biochemistry. Understanding the underpinnings of gene expression have been my enduring interest;
thus I transitioned from working on the differentiating bacterium, Caulobacter crescentus, in the Shapiro lab, to the
differentiating protozoan parasite, Trypanosome brucei. My post doctoral work in Dr. George Cross' laboratory of
Molecular Parasitology at the Rockefeller was the foundation for my independent work on gene expression in trypanosomes.
My laboratory at New Jersey Medical School has uncovered several of the molecular components of specific RNA polymerase
II-dependent genes that reside in the cell nucleus and specific protein-mRNA interactions that function in the cell
cytoplasm. Our goals are to develop therapies for the amelioration of neglected diseases that include trypanosomes and
related parasites, and to discover key pieces of the genetic regulatory networks that underlie biology.

Education

PHD, 1984, Albert Einstein College of Medicine of Yeshiva University
B.S., 1977, Douglass College/Rutgers University

Relevant Publications

Liu W, Das A, Morales R, Banday M, Aris V, Lukac DM, Soteropoulos P, Wah DA, Palenchar J, Bellofatto V., Chromatin immunoprecipitation and microarray analysis reveal that TFIIB occupies the SL RNA gene promoter region in Trypanosoma brucei chromosomes. Mol Biochem Parasitol. 2012 Dec;186(2):139-42. doi: 10.1016/j.molbiopara.2012.09.003. Epub 2012 Sep 19. PMID: 22999857

Das A, Morales R, Banday M, Garcia S, Hao L, Cross GA, Estevez AM, Bellofatto V. The essential polysome-associated RNA-binding protein RBP42 targets mRNAs involved in Trypanosoma brucei energy metabolism. RNA. 2012 Nov;18(11):1968-83. doi: 10.1261/rna.033829.112. Epub 2012 Sep 10. PMID: 22966087

Utter, C. J., Garcia, S. A., Milone, J., & Bellofatto, V. (2011). Poly (A)-specific Ribonuclease (PARN-1) function in stage-specific mRNA turnover in Trypanosoma brucei. Eukaryot Cell. 2011 Sep;10(9):1230-40. doi: 10.1128/EC.05097-11. Epub 2011 Jul 8. PMID: 21743004

Das, A., & Bellofatto, V. (2009). The non-canonical CTD of RNAP-II is essential for productive RNA synthesis in Trypanosoma brucei. PLoS One. 2009 Sep 9;4(9):e6959. doi: 10.1371/journal.pone.0006959. PMID: 19742309

Ibrahim BS, Kanneganti N, Rieckhof GE, Das A, Laurents DV, Palenchar JB, Bellofatto V, Wah DA. (2009). Structure of the C-terminal domain of transcription factor IIB from Trypanosoma brucei. Proceedings of the National Academy of Sciences, 106(32), 13242-13247. PMID: 19666603

Banerjee H, Palenchar JB, Lukaszewicz M, Bojarska E, Stepinski J, Jemielity J, Guranowski A, Ng S, Wah DA, Darzynkiewicz E, Bellofatto V. (2009). Identification of the HIT-45 protein from Trypanosoma brucei as an FHIT protein/dinucleoside triphosphatase: Substrate specificity studies on the recombinant and endogenous proteins.RNA,15(8), 1554-1564. PMID: 19541768

Das, A., Banday, M., & Bellofatto, V. (2008). RNA polymerase transcription machinery in trypanosomes. Eukaryotic cell, 7(3), 429-434. PMID: 17951525

Areas Of Interest

Trypanosomes

Gene expression

Infectious Disease

RNA binding proteins

RNA polymerase II

mRNA

Regulation of Gene Expression in Parasitic Protozoa

The quality of life of millions is compromised by parasitic infection. Trypanosomes, spread by
tsetse flies to humans and grazing animals in sub- Saharan Africa or by reduviid bugs in many
areas of South and Central America, cause debilitating and lethal disease. These unicellular,
eukaryotic organisms reside extracellularly, in the bloodstream and tissue spaces of the host or
intracellularly, within smooth muscle cells. As parasites, they readily differentiate during
their complex life cycle to adapt to numerous nutritional conditions and immunological
challenges. Scientific research has uncovered a multitude of unusual genetic, biochemical and
structural properties of trypanosomes. We hope that by exploiting some of these parasite-specific
traits we can devise ways to alleviate disease.



Messenger RNA molecules in eukaryotic cells usually begin as large precursor forms that are cut
and spliced to produce stable and translatable mature mRNAs. In most cases, a single molecule of
RNA is the starting material. However, trypanosomes synthesize messenger RNAs in a unique way.
All trypanosome mRNAs are produced by a splicing reaction that fuses a short, capped RNA (called
the spliced leader) to each protein coding pre-mRNA. The spliced leader is essential for gene
expression; however, its synthesis, regulation and role in mRNA maturation and translation remain
enigmas. Our work is focused on understanding the nucleic acid and protein signals responsible
for these processes.



To dissect the components of the spliced leader gene essential for its transcription, we are
using a combination of genetic analysis and DNA-protein binding studies. Protein purification and
subsequent genetic analysis will enable us to identify unique trans-acting factors of the spliced
leader transcription pathway. This work has lead to our discovery of PBP-1, a basal transcription
factor that is an essential component of the spliced leader RNA gene expression pathway.



Since the 5' cap present on each mature mRNA is added in trans in trypanosomes, the "one pre-mRN
-one mRNA" rule observed in other eukaryotes does not apply here. Thus, trypanosomes synthesize
polycistronic pre-mRNAs that are cleaved into multiple mature mRNAs via RNA trans- splicing.
Post- transcriptional regulation of gene expression is probably orchestrated in part by
differential spliced leader addition to specific sites within polycistronic pre-mRNAs. We are
investigating the signals that direct the spliced leader to specific precursor transcripts whose
mRNA levels appear to be post-transcriptionally regulated.



Finally, we have developed an in vitro RNA turnover system that is enabling us to explore and
define the signals that regulate differential mRNA turnover as a function of life cycle stage in
trypanosomes.