Maintenance Scientist

Jayaraju Dheekollu, PhD

*** ******* *** *****: 215-***-****

Drexel Hill, PA 19026 Email: ***********@*****.***

Summary of Qualifications

Molecular Virologist with eighteen years of experience

Track record of high-impact publications

Knowledge of state-of-the-art virology, immunology, molecular biology and cell biology methods

Experience in sterile technique and safe handling of infectious BSL2 organisms

Expert in the design and implementation of in vitro and in vivo experimentation and assays

Up-to-date knowledge of antiviral treatment modalities

Education and Training

Postdoctoral Fellowship, Wistar Institute, Philadelphia, PA 2003-2005

Postdoctoral Fellowship, Hôtel-Dieu de Québec, Laval University, Quebec, Canada 2000-2002

PhD, Biochemistry, University of Hyderabad, Hyderabad, Telangana 500046, India 1993-1999

M.Sc, Biochemistry, University of Hyderabad, Hyderabad, Telangana 500046, India 1991-1993

B.Sc, Biology, Nagarguna University, Guntur, Andhra Pradesh 522510, India 1987-1991

Research Experience

Wistar Institute, Philadelphia, PA 2005-2021

Scientist

To understand the molecular mechanisms of EBNA1 and LANA role in

gamma herpes latent viral genome replication and maintenance.

Identification of Host Replication and recombination factors involved in the maintenance mechanism of EBV and KSHV Genomes during Latency.

Study for Epigenetic Regulation of EBV Latent Origin of replication (OriP).

Center for breakthrough medicine, King of Prussia, PA 2022-Present

Scientist

TCID50 Analytical method development and qualification for GMP use.

Developing Mycoplasma test method.

Developing the assays for detecting Replication competent lentiviral vectors (RCL) and Adeno-associated viral Vectors (rcAAV)

Potency assay for Gene Therapy drug products.

Technical Skills

Cell Culture

Experience in culturing primary cells including human T-cells and B-cells from blood and spleen,

Extensive experience culturing adherent and non-adherent mammalian cell lines including: 293T, HeLa, Raji, Mutu1, LCLs, BCBL1, BC1, BC3, BJAB, DJ75, Hct116, M81, ATLD1, ATLD3, GM15808, GM 07078 and U20S

Molecular Biology

Cloning

Site-directed mutagenesis

DNA and RNA extraction and sequencing

RT-PCR

qPCR

ddPCR

Microarray

Western blot

Transfection

Reporter assays

Microscopy (fluorescent and confocal)

Lentiviral transduction

CHIP

CHIP Sequence

Southern blot

2d gel analysis of replication intermediates

Protein Gel shifts

Plasmid replication and maintenances assays

Protein purification

PFGF electrophoresis

DNA sequencing Gels

CRISPR technology

Immunology

Western blot

IFA

ELISA development

Immunohistochemistry

Proliferation assays

Flow cytometry

Virology

Experience with BSL2 and BSL3 agents

Viral infectivity

Viral genome titer

Viral potency

Viral-cell fusion assay

Virus purification

B cell Immortalization assays

Tissue culture (cell lines and primary cells)

EBV Bac mutagenesis

Have worked with the following viruses: Herpesviruses (EBV, KSHV, AD, AAV, Lenti)

Academic Highlights

1993-1995 Junior Research Fellowship JRF, Council of scientific and industrial research (CSIR), India

1995-1999 Senior Research Fellowship SRF, Council of scientific and industrial research (CSIR), India

Publications

Dheekollu J, Wiedmer A, Soldan S and Lieberman PM.

Post-translational regulation of EBNA 1 by Lysine Hydroxylase Plod1

In review

Mei Y, Messick TE, Dheekollu J, Kim HJ, Molugu S, Muñoz LJC, Moiskeenkova-Bell V, Murakami K, Lieberman PM. Cryo-EM Structure and Functional Studies of EBNA1 Binding to the Family of Repeats and Dyad Symmetry Elements of Epstein-Barr Virus oriP. J Virol. 2022 Aug 29

Dheekollu J, Wiedmer A, Kasirajan A, Deakyne JS, Messick TE, and Lieberman PM. Cell Cycle-Dependent EBNA1-DNA Cross-Linking Promotes Replication Termination at oriP and Viral Episome Maintenance. Cell. 2021 Feb 4;184(3):643-65

Li F, Deng Z, Zhang L, Wu C, Jin Y, Hwang I, Vladimirova O, Xu L, Yang L, Lu B, Dheekollu J, Li JY, Feng H, Hu J, Vakoc CR, Ying H, Paik J, Lieberman PM, Zheng H. ATRX loss induces telomere dysfunction and necessitates induction of alternative lengthening of telomeres during human cell immortlization. EMBO J. 2019 Oct 1;38(19)

De Leo A, Deng Z, Vladimirova O, Chen HS, Dheekollu J, Calderon A, Myers KA, Hayden J, Keeney F, Kaufer BB, Yuan Y, Robertson E, Lieberman PM. LANA oligomeric architecture is essential for KSHV nuclear body formation and viral genome maintenance during latency. PLoS Pathog. 2019 Jan 25;15(1).

Malecka KA, Dheekollu J, Deakyne JS, Wiedmer A, Ramirez UD, Lieberman PM, Messick TE. Structural Basis for Cooperative Binding of EBNA1 to the Epstein-Barr Virus Dyad Symmetry Minimal Origin of Replication. J Virol. 2019 Sep 30;93(20

Dheekollu J, Malecka K, Wiedmer A, Delecluse HJ, Chiang AKS, Altieri DC, Messick TE, Lieberman PM. Carcinoma-risk variant of EBNA1 deregulates Epstein-Barr Virus episomal latency. Oncotarget. 2019 Jul 8;10(43):4505-4506.

Deng Z, Kim ET, Vladimirova O, Dheekollu J, Wang Z, Newhart A, Liu D, Myers JL, Hensley SE, Moffat J, Janicki SM, Fraser NW, Knipe DM, Weitzman MD, Lieberman PM. HSV-1 remodels host telomeres to facilitate viral replication. Cell Rep. 2014 Dec 24;9(6):2263-78.

Dheekollu J, Wiedmer A, Hayden J, Speicher D, Gotter AL, Yen T, Lieberman PM. Timeless links replication termination to mitotic kinase activation. PLoS One. 2011 May 6;6(5).

Dheekollu J, Wiedmer A, Sentana-Lledo D, Cassel J, Messick T, Lieberman PM. HCF1 and OCT2 Cooperate with EBNA1 To Enhance OriP-Dependent Transcription and Episome Maintenance of Latent Epstein-Barr Virus. J Virol. 2016 May 12;90(11):5353-5367.

Tsai K, Chan L, Gibeault R, Conn K, Dheekollu J, Domsic J, Marmorstein R, Schang LM, Lieberman PM. Viral reprogramming of the Daxx histone H3.3 chaperone during early Epstein-Barr virus infection. J Virol. 2014 Dec;88(24):14350-63.

Deng Z, Glousker G, Molczan A, Fox AJ, Lamm N, Dheekollu J, Weizman OE, Schertzer M, Wang Z, Vladimirova O, Schug J, Aker M, Londoño-Vallejo A, Kaestner KH, Lieberman PM, Tzfati Y. Inherited mutations in the helicase RTEL1 cause telomere dysfunction and Hoyeraal-Hreidarsson syndrome. Proc Natl Acad Sci U S A. 2013 Sep 3;110(36).

Dheekollu J, Chen HS, Kaye KM, Lieberman PM. Timeless-dependent DNA replication-coupled recombination promotes Kaposi's Sarcoma-associated herpesvirus episome maintenance and terminal repeat stability. J Virol. 2013 Apr;87(7):3699-709.

Kumala S, Fujarewicz K, Dheekollu J, Rzeszowska-Wolny J, Hancock R. Repair of DNA strand breaks in a minichromosome in vivo: kinetics, modeling, and effects of inhibitors. PLoS One. 2013;8(1)

Leman AR, Dheekollu J, Deng Z, Lee SW, Das MM, Lieberman PM, Noguchi E. Timeless preserves telomere length by promoting efficient DNA replication through human telomeres. Cell Cycle. 2012 Jun 15;11(12):2337-47.

Tempera I, Wiedmer A, Dheekollu J, Lieberman PM. CTCF prevents the epigenetic drift of EBV latency promoter Qp. PLoS Pathog. 2010 Aug 12;6(8):e1001048.

Dheekollu J, Lieberman PM. The replisome pausing factor Timeless is required for episomal maintenance of latent Epstein-Barr virus. J Virol. 2011 Jun;85(12):5853-63.

Deng Z, Dheekollu J, Broccoli D, Dutta A, Lieberman PM. The origin recognition complex localizes to telomere repeats and prevents telomere-circle formation. Curr Biol. 2007 Nov 20;17(22):1989-95.

Dheekollu J, Deng Z, Wiedmer A, Weitzman MD, Lieberman PM. A role for MRE11, NBS1, and recombination junctions in replication and stable maintenance of EBV episomes. PLoS One. 2007 Dec 5;2(12).

Wang P, Day L, Dheekollu J, Lieberman PM. A Redox-sensitive Cysteine in Zta is required for Epstein-Barr virus lytic cycle DNA replication. J Virol. 2005 Nov;79(21):13298-309.

Dheekollu j, Kondapi AK. Anti-cancer copper salicylaldoxime complex inhibits topoisomerase ii catalytic activity. Current science, vol. 81, no. 7 (10 october 2001), pp. 787-792

Vashisht Gopal YN, Dheekollu J, Kondapi AK. Topoisomerase II is a cellular target for antiproliferative cobalt salicylaldoxime complex. Arch Biochem Biophys. 2000 Apr 1;376(1):229-35. doi: 10.1006/abbi.1999.1692.

Dheekollu J, Gopal YN, Kondapi AK. Topoisomerase II is a cellular target for antiproliferative cobalt salicylaldoxime complex. Arch Biochem Biophys. 1999 Sep 1;369(1):68-77.

Gopal YN, Dheekollu J, Kondapi AK. Inhibition of topoisomerase II catalytic activity by two ruthenium compounds: a ligand-dependent mode of action. Biochemistry. 1999 Apr 6;38(14):4382-8.

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