Quality Control Assistant

Venkateswarlu. Popuri, PhD

Dept of Microbiology and Molecular Genetics,

Ph : 001-240-***-****

University of California, Davis.

E.mail: ******@*******.***; ******.*************@*****.***

Summary: High potential Research scientist studying DNA damage signaling and DNA repair

pathways. Proficiency in in-vitro biochemistry with recombinant protein purification and all basic

biochemical, molecular biology and cloning techniques along with a good experience in cell culture and

confocal and fluorescent live microscopy. Good exposure in pharmaceutical formulations & QC. Able to

work independently, and also an exceptional team player. Well organized, focused and a very good

communicator. Good experience in writing research articles / publications.

Education:

2004 – 2008: PhD: International Centre for Genetic engineering & Biotechnology

(ICGEB), Trieste, Italy.

2002 – 2003: Master’s in Biotechnology. (M.Tech). Jawaharlal Nehru Technological

University, Hyderabad, India.

1997 – 2001: Bachelor’s in Pharmacy. (B.Pharm). Sri Venkateswara college of

pharmacy, Osmania University, Hyderabad.

Professional Experience:

Jan 2014 – Present: Assistant Project Scientist III at University of California, Davis.

Research summary: Studying telomere maintenance mechanisms in cancer cells. Telomeres are ends of

chromosomes and telomere shortening occurs after every cell division cycle resulting in senescence and

aging processes. Cancer cells, on the other hand, maintain telomere length for several replication cycles. In

specific, we are currently studying molecular mechanisms such as ALT pathways of telomere maintenance

in cancer cells. ALT cells have somatic mutations in ATRX. Loss of ATRX, an SF2-family chromatin

remodeling protein, leads to X-linked alpha thalassemia mental retardation syndrome. We are currently

investigating potential functional role for ATRX in telomere replication. Loss of ATRX is a hall mark

feature of ALT cells and we are currently investigating why loss of ATRX would lead to a ALT phenotype.

Jan 2009 – Jan 2014: Post doctoral research associate, at National Institute of Aging,

(NIA / NIH), Baltimore, MD.

Supervisor: Dr. Vilhelm A. Bohr, Cheif, LMG, NIA/NIH.

Research summary: RecQ helicases are care takers of human genome and are termed as tumor suppressors

and play important roles in maintenance of genomic stability. We have elucidated potential functions of

RECQL5 and RECQL1 in DNA replication that are recently published in peer reviewed journals. We have

observed for the first time, a synthetic lethal interaction among these human RecQ helicases. RECQL5

provides strong potential back up functions to WRN syndrome helicase and RECQ L5 is essential for cell

survival in the absence of WRN as depletion of RECQL5 severely compromises DNA replication and

causes cell death in the absence of WRN. Further, we deciphered the role of RECQL1, a potential tumor

suppressor, at stalled replication forks and its eventual role in replication fork progression, DNA damage

signaling pathways and genomic stability. Depletion of RECQL1 induces hyper phosphorylation of RPA

and activation of p-CHK1, indicating replicative stress. Additionally, we recently reported a novel role for

RECQL1 in telomere maintenance. RECQL1 can unwind various secondary structures associated with

telomeres like that of Telomeric D-loops and Holliday junctions. Stable depletion of RECQL1 leads to

telomere shortening, aphidicolin-induced telomere fragility and elevated telomere sister chromatid

exchanges.

Oct 2004 – Dec. 2008: PhD on the “Structural, functional and enzymatic characterization

of human RecQ1 helicase”, under the supervision of Dr. Alessandro Vindigni, ICGEB,

Trieste, Italy.

Supervisor: Dr. Alessandro Vindigni.

Research summary: Performed peer reviewed research, in elucidating the structural, functional and

enzymatic analysis of human RecQL1 helicase and further elucidating the substrate specificity in

comparison to that of other human RecQ helicases. Gel filtration experiments supported by Electron

microscopy and single turn over unwinding kinetics suggest that RECQL1 unwinds DNA as a monomer.

Oct 2003 – Sep 2004: Research Assistant at Center for Cellular and Molecular Biology,

(CCMB), Hyderabad, India.

Supervisor: Dr. N. Madhusudhan Rao.

Research summary: Worked as a Research assistant on characterization and cloning of a gene encoding for

a fungal protease. RNA was isolated from a fungal species, and cDNA library was constructed by RT-PCR

and subsequent cloning of the fungal protease gene. Sponsored by CSIR-NMITLI, proteases are very

helpful for tanning in the leather industry.

Jan 2003 – July 2003: Research trainee at National Institute of Immunology, (NII), New

Delhi, India.

Supervisor: Dr. Amulya K. Panda

Research summary: Purification of recombinant Polyketide synthase from the inclusion bodies of E.Coli.

Performed isolation and solubilization of inclusion bodies from the bacterial expression systems. In vitro

refolding by pulsatile dilution to form a functional and folded form.

Apr 2001: Research Intern in Rexer Pharma, Hyderabad, on Quality control of Drug

dosage forms.

Worked as a Research Intern (B. Pharm) on Manufacturing & Quality Control of various dosage forms.

Good Exposure in the manufacturing of solid drug dosage forms, following GMP. Good exposure in the

Quality control, Potency assays, Dissolution & Disintegration studies, Content uniformity. & All QC

assays.

Technical skills:

● Recombinant protein expression and purification in E.coli and baculo-virus systems. Good experience in

isolation and solubilization of inclusion bodies & subsequent protein folding.

● Protein stability studies with an understanding of steady-state and pre-steady state enzyme kinetics and

Proficiency in setting up radiometric and florescent enzymatic assays.

● FRET, FACS, Gel Electrophoresis & all basic Biochemistry, Molecular Biology & Microbiology

techniques like CHIP, ELISA, PCR, cloning, restriction digestion analysis, transformation etc…

● Good Experience in Cell Culture and all types of Cell based assays. Usage of Lentiviral shRNA to

generate stable knockdown cells, RNA isolation techniques, RT-PCR.

Confocal Microscopy and/or live cell fluorescent microscopy, Use of laser to induce double strand

breaks.

● Teloblot, Telomere Q-FISH and Co-FISH experiments.

● SDS-PAGE / Southern / Western Analysis and Co-immunoprecipation assays. Immunofluorescence and

Confocal Microscopy.

● Good Exposure in GLP, GMP’s, SOP’s, Pharmaceutical formulations, QC & drug discovery.

● Excellent ongoing experience in DNA rep lication, recombination and repair mechanisms. Proficiency in

DNA repair pathways and oncology mechanisms.

● Exposure and understanding of Pharmacological / cell toxicity mechanisms.

Publications:

Popuri V, Bachrati CZ, Muzzolini L, Mosedale G, Hickson ID, Vindigni A. The

human RecQ helicases, BLM and RECQ1, display distinct DNA substrate

specificities. 2008; J. Biol. Chem, 283, 177**-*****.

Popuri V, Jing H, Ramamoorthy M, Croteau DL and Bohr VA. RECQL5 plays

both co-operative and complementary roles with that of WRN syndrome helicase.

2012; Nucleic Acids Res., 41-881-99.

Popuri V, Hsu J, Khadka P, Horvath K, Liu Y, Croteau DL, and Bohr VA.

Human RECQL1 participates in telomere maintenance. 2014; Nucleic Acids

Res., (Epub ahead of print).

Popuri V, Croteau, DL, and Bohr VA. Substrate specific stimulation of NEIL1

by WRN but not other RecQ helicases. 2010; DNA Repair, 9, 636-642.

Popuri V, Ramamoorthy M, Tadokoro T, Singh DK, Croteau, DL and Bohr

VA. Recruitment and retention dynamics of RECQL5 at DNA double strand break

sites. 2012; DNA Repair, 11, 624-635.

Popuri V, Croteau DL, Brosh RM Jr and Bohr VA. RECQ1 is required for

cellular resistance to replication stress and catalyzes strand exchange on stalled

replication fork structures. 2012; Cell Cycle, 11, 4252-65.

Popuri V, Ferrarelli LK, Ghosh AK, Tadokoro T, Canugovi C, Hsu JK, Croteau

DL and Bohr VA. The RECQL4 protein, deficient in Rothmund-Thomson

syndrome is active on telomeric D-Loops containing DNA metabolism blocking

lesions. 2013; DNA repair, 12, 518-528.

Singh DK, Popuri V, Kulikowicz T, Shevelev I, Ghosh AK, Ramamoorthy M,

Rossi ML, Janscak P, Croteau DL, Bohr VA. The human RecQ helicases BLM

and RECQL4 cooperate to preserve genome stability. 2012; Nucleic Acids Res.,

40, 6632-648.

Tadokoro T, Ramamoorthy M, Popuri V, Sykora P, Rybanska I, Croteau DL

and Bohr VA. Human RECQL5 participates in removal of endogenous DNA

damage. 2012; Mol Biol Cell., 23, 4273-85.

Ramamoorthy M, May A, Tadokoro T, Popuri V, Seidman, MM, Croteau, DL

and Bohr VA. The RecQ helicase RECQL5 participates in psoralen induced

interstrand cross-link repair. 2013; Carcinogenesis, 34, 2218-2230.

Pike AC, Shrestha B, Popuri V, Burgess-Brown N, Muzzolini L, Costantini S,

Vindigni A, Gileadi O. Structure of the human RECQ1 helicase reveals a putative

strand-separation pin. 2009; Proc.Natl Acad Sci U S A (PNAS), 106, 1039-44.

Muzzolini L, Beuron F, Patwardhan A, Popuri V, Cui S, Niccolini B, Rappas

M, Freemont Ps, Vindigni A. Different quaternary structures of human RECQ1

are associated with its dual enzymatic activity. 2007; PLoS Biol., 5,e20.

Aamann MD, Hvitby C, Popuri V, Muftuoglu M, Lemminger L, Skeby CK,

Keijzers G, Ahn B, Bjørås M, and Bohr VA, Stevnsner T. Cockayne Syndrome

group B protein stimulates NEIL2 DNA glycosylase activity. 2014; Mech Aging

Dev., 135, 1-14.

Reviews:

Popuri V, Tadokoro T, Croteau DL and Bohr VA. Human RECQL5: Guarding

the crossroads of DNA replication and transcription and providing backup

capability. 2013; Critical Reviews in Biochemistry and Molecular Biology, 48,

289-299.

Croteau DL, Popuri V, Opresko PL and Bohr VA. Human RecQ Helicases in

DNA Repair, Recombination, and Replication. 2014; Ann Rev Biochem., 83,

(Epub ahead of print).

Academic merits & other achievements:

Awarded NIH FARE Award for best research abstract in 2011.

Presented a poster at prestigious Gordon Research Conferences 2012: DNA

damage, mutation and cancer.

Presented a poster at ASBMB, Genome replication and repair symposium,

2013.

Reviewed over 10 publications for peer reviewed journals like Cancer research,

PLoS One, DNA repair, International journal of Biochemistry and cell biology,

Mechanisms of Aging and Development, Current Aging Science, Current

Molecular medicine.

Presented Posters at BARS symposium and NIH retreat.

Qualified in National level GATE Scholarships (2001) with 90.13 percentile.

Topper in M.Tech (Bio technology).

Secured a Gold medal for being the topper in Bachelor’s in Pharmacy.

References:

Dr. Vilhelm Bohr,

Chief, LMG,

NIA / NIH,

Baltimore, MD 21224

E-mail: *****@****.***.***

Dr. Alessandro Vindigni, Dr. S. Rama Krishna,

Associate Professor, Scientist, Pharmacology Division,

Saint Louis School of medicine, Indian Institute of Chemical Technology,

Saint Louis, MO 63104. Tarnaka, Hyderabad.

E-mail: ********@***.*** E-mail: ******@****.***.**

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