Drohat, Alexander | University of Maryland School of Medicine

Academic Title:

Professor

Primary Appointment:

Biochemistry and Molecular Biology

Email:

adrohat@som.umaryland.edu

Location:

108 N. Greene St., 419

Education and Training

University of Maryland, BS, Aerospace Engineering

University of Maryland School of Medicine, PhD, Biochemistry and Molecular Biology

Johns Hopkins University School of Medicine, NIST and Center for Advanced Research in Biotechnology, Postdoctoral Fellow

Biosketch

Our research centers on two broad areas, DNA repair and epigenetic regulation through DNA methylation. The nucleobases of DNA are amenable to a broad range of chemical alterations, a feature that enables enzyme-mediated modifications but also allows for threatening DNA damage. We study enzymes that find and repair DNA lesions, thereby maintaining genomic integrity and protecting against cancer and other diseases. We also investigate enzymes that perform essential functions in epigenetic regulation, by acting on modified DNA bases. We use a broad range of biochemical, biophysical, structural, and molecular approaches, and collaborate with many other research groups. Some of our current research interests are summarized below.

Research/Clinical Keywords

DNA Repair, Epigenetics and DNA methylation, Enzymology, NMR, Structural Biology, SUMO modification

Highlighted Publications

Pidugu LS, Servius HW, Espinosa KB, Cook ME, Varney KM, Drohat AC (2024) Sumoylation of thymine DNA glycosylase impairs productive binding to substrate sites in DNA, J Biol Chem 300 (11), 107902.

Pidugu LS, Servius HW, Sevdalis SE, Cook ME, Varney KM, Pozharski E, Drohat AC (2023) Characterizing inhibitors of human AP endonuclease 1. PLoS ONE 18(1): e0280526 (PMCID: PMC9847973).

Servius HW, Pidugu LS, Sherman ME, Drohat AC (2023) Rapid Excision of Oxidized Adenine by Thymine DNA Glycosylase, J Biol Chem 299:102756.

Pidugu LS, Bright H, Lin WJ, Majumdar C, Van Ostrand RP, David SS, Pozharski E, Drohat AC (2021) Structural Insights into the Mechanism of Base Excision by MBD4, J Mol Biol 433, 167097.

Pidugu LS, Dai Q, Malik SS, Pozharski E, Drohat AC (2019)Excision of 5-carboxylcytosine by Thymine DNA Glycosylase,J Am Chem Soc141, 18851-61.

Dow BJ, Malik SS, Drohat AC (2019) Defining the Role of Nucleotide Flipping in Enzyme Specificity Using ¹⁹F NMR, J Am Chem Soc 141, 4952-62.

Complete list of publications: NCBI and Google Scholar

DNA Repair

Often termed the “5th base” in DNA, 5-methylcytosine (mC) is a key epigenetic mark in eukaryotes, and it functions in restriction modification systems of archaea and bacteria. However, mC also threatens genetic and epigenetic integrity. Deamination of mC to T generates G/T mispairs, and, upon replication, CàT transitions. Through this process, mC deamination causes many point mutations in cancer and genetic disease. Two human glycosylases remove T from G/T mispairs, TDG (thymine DNA glycosylase) and MBD4 (methyl binding domain IV). Like other glycosylases, they flip a damaged base into their active site and cleave the base-sugar bond; follow-on base excision repair (BER) enzymes complete the repair process. While most glycosylases excise bases that are foreign to DNA (e.g., uracil), these mismatch enzymes remove a canonical base, thymine, from rare G/T mispairs but not from the vast background of A:T pairs. Because aberrant action on A:T pairs can be mutagenic and cytotoxic, specificity is critical for these enzymes, and we are studying how it is attained. We also study how TDG and MBD4 recognize other damaged bases, including uracil and 5-fluorouracil (5FU), among others. TDG excision of 5FU contributes to the antitumor effects of 5FU, which is used to treat cancer.

Servius et al (2023) J Biol Chem

Pidugu et al. (2021) J Mol Biol

Dow et al (2019) J Am Chem Soc

Coey et al (2016) Nucleic Acids Res

Maiti et al. (2012) Proc Natl Acad Sci

Epigenetic Regulation - Base Excision Repair in active DNA Demethylation

Methyltransferases are known to convert C to mC, but the process for reversing this epigenetic modification had remained unclear. One mechanism involves DNA replication without subsequent remethylation. Recent studies establish a pathway for active DNA demethylation, involving TET (ten-eleven translocation) enzymes and TDG-initiated BER (Fig. 1). TET enzymes catalyze stepwise oxidation of mC, to give 5-hydroxymethyl-C (hmC), then 5-formyl-C (fC), and 5-carboxyl-C (caC). TDG excises fC and caC, and subsequent BER steps restore cytosine. This central function in epigenetic regulation likely explains findings that TDG is essential for embryonic development. We study the role of TDG and other BER enzymes in active DNA demethylation.

Pidugu et al (2019) J Am Chem Soc

Pidugu et al (2016) Biochemistry

Maiti et al (2013) J Am Chem Soc

Maiti and Drohat (2011) J Biol Chem

Protein Regulation by SUMO binding and SUMO conjugation

TDG can be covalently modified by SUMO (small ubiquitin-like modifier) proteins, and it also features a SUMO interacting motif (SIM) that binds non-covalentlyto SUMO domains. The SIM can bind free SUMO, a SUMO domain that is tethered to another protein, and intramolecularSUMO (tethered to TDG). We are investigating the effect of SUMO binding and SUMO conjugation on TDG function(s), and testing the current paradigm that SUMO modification of product-bound TDG is needed to relieve product inhibition and allow for efficient catalytic turnover. We are also studying how SUMO modification of TDG is regulated in cells.

Pidugu et al (2024) J Biol Chem

Coey and Drohat (2018) Nucleic Acids Res

McLaughlin et al (2016) J Biol Chem

Coey at al. (2014) J Biol Chem

2022 - Elected Fellow, American Association for the Advancement of Science

2020-2025 - R35 MIRA Award, National Institute General Medical Sciences

2018-2022 - Member, NIH Molecular Genetics A (MGA) study section

2011-2015 - Member, DNA Mechanism in Cancer Peer Review Committee, American Cancer Society

2011-2014 - Editorial Advisory Board, ASBMB Today

2001 - Travel Award, American Chemical Society National Meeting

1998-2000 - National Research Council Postdoctoral Research Fellowship

1997 - Graduate Student Research Day Award, University of Maryland Baltimore