About

Professor Rob McGinty, M.D., Ph.D., is the Principal Investigator of the McGinty Lab at the University of North Carolina at Chapel Hill. He holds a Ph.D. from The Rockefeller University and an M.D. from Weill Cornell Medical College. His research focuses on epigenetic signaling at atomic resolution, exploring the molecular mechanisms underlying epigenetic regulation. The lab's work involves detailed structural and biochemical studies to understand how epigenetic modifications influence gene expression and cellular function, contributing to the broader understanding of epigenetic processes in health and disease.

Research topics

• Biology
• Cell biology
• Computational biology
• Biophysics
• Genetics
• Chemistry
• Biochemistry

Selected publications

• PELP1 coordinates the modular assembly and enzymatic activity of the rixosome complex

  Science Advances · 2025-07-25 · 2 citations

  articleOpen access

  The rixosome is a large multisubunit complex that initiates RNA decay during critical nuclear transactions including ribosome assembly and heterochromatin maintenance. The overall architecture of the complex remains undefined because several subunits contain intrinsically disordered regions (IDRs). Here, we combined structural and functional approaches to establish PELP1 as the central scaffold of the rixosome upon which the enzymatic subunits modularly assemble. The C-terminal half of PELP1 is composed of a proline-rich IDR that mediates association with the AAA-ATPase MDN1, histones, and the SUMO-specific protease SENP3. The PELP1 IDR contains a glutamic acid-rich region that we establish can chaperone the histone octamer in vitro. Last, the x-ray structure of a small linear motif (SLiM) from the PELP IDR bound to SENP3 reveals how PELP1 allosterically activates SUMO protease activity. This work provides an integrated structural model for understanding the rixosome's dynamic architecture and how it modularly coordinates several cellular functions.

  PublisherDOI

• An intrinsically disordered region of histone demethylase KDM5A activates catalysis through interactions with the nucleosomal acidic patch and DNA

  bioRxiv (Cold Spring Harbor Laboratory) · 2025-05-02

  preprintOpen access

  ABSTRACT Lysine demethylase 5A (KDM5A) plays a key role in the regulation of chromatin accessibility by catalyzing the removal of trimethyl marks on histone H3K4 (H3K4me3). KDM5A is also an oncogenic driver, with overexpression of KDM5A observed in various cancers, including breast, lung, and ovarian cancer. Past studies have characterized the functions of KDM5A domains, including KDM5A interactions with the histone H3 tail, but have yet to identify the broader mechanisms that drive KDM5A binding to the nucleosome. Through investigation of binding and catalysis on nucleosome substrates, we uncovered multivalent interactions of KDM5A with the H2A/H2B acidic patch and DNA that play crucial roles in the regulation of catalytic activity. We also identified an intrinsically disordered region (IDR) containing bifunctional arginine-rich motifs capable of binding to both the histone H2A/H2B acidic patch and nucleosomal DNA that is necessary for catalysis on nucleosome substrates. Our findings both elucidate previously unknown mechanisms that regulate KDM5A catalytic activity and reveal the ability of an IDR to engage in multiple interactions with chromatin. ARTICLE HIGHLIGHTS The intrinsically disordered region of KDM5A binds the acidic patch and DNA. Interactions with the nucleosome are mediated by arginine-rich motifs in the IDR. The IDR properly orients KDM5A on the nucleosome to enable catalysis.

  PublisherOA PDFDOI

• Mechanistic Insights into the Stimulation of Dot1L-Mediated Methylation of Histone H3 by Semisynthetically Ubiquitylated Histone H2b

  Digital Commons - RU (Rockefeller University) · 2025-09-08

  articleOpen access1st authorCorresponding

  Post-translational modification of histones plays an integral role in regulation of chromatin-templated processes through modulation of chromatin structure and function. One such modification, ubiquitylation of histone H2B on lysine 120 (uH2B), has been correlated with enhanced methylation of lysine 79 (K79) of histone H3 by K79-specific methyltransferase, disruptor of telomeric silencing-like (Dot1L/KMT4). However, the specific function of uH2B in this crosstalk pathway was not understood, in part due to the challenges associated with isolating or generating homogeneously ubiquitylated H2B for use in biochemical studies. As both modifications are integral to transcriptional regulation and DNA damage repair, full elucidation of their functions is critical to understanding their roles in development and disease. In this thesis, a chemical strategy is presented for the preparation of native uH2B. Two traceless orthogonal expressed protein ligation (EPL) reactions were used for this purpose, one employing a photolytically removable ligation auxiliary, and the other, a cysteine-mediated ligation followed by a desulfurization to restore the native sequence. Reconstitution of semisynthetic uH2B into chemically defined nucleosomes, followed by biochemical analysis, revealed a direct role for uH2B in the stimulation of Dot1L-mediated methylation of H3K79. Although recruitment of Dot1L to the nucleosomal surface by uH2B could be excluded, comprehensive mechanistic analysis was precluded by systematic limitations in the ability to generate native uH2B in large-scale. To overcome this shortcoming, a highly optimized synthesis of ubiquitylated H2B bearing a Gly76Ala point mutation (uH2BG76A) was developed, yielding tens of milligrams of ubiquitylated protein. This mutant was indistinguishable from native uH2B by Dot1L, allowing for detailed studies of the resultant trans-histone crosstalk pathway. Kinetic and structure activity relationship analyses using uH2BG76A suggest a non-canonical role for ubiquitin in the enhancement of the chemical step of H3K79 methylation. This enhancement likely results from an allosteric change in the nucleosome and/or Dot1L following H2B ubiquitylation. Current work is aimed at further elucidation of the molecular mechanism of uH2B-mediated stimulation of Dot1L and the role of uH2B in other chromatin templated-processes.

  PublisherOA PDFDOI

• Nickel-NTA lipid-monolayer affinity grids allow for high-resolution structure determination by cryo-EM

  bioRxiv (Cold Spring Harbor Laboratory) · 2025-04-24

  preprintOpen access

  Grid preparation is a rate-limiting step in determining high-resolution structures by single particle cryogenic electron microscopy (cryo-EM). Particle interaction with the air-water interface often leads to denaturation, aggregation, or a preferred orientation within the ice. Some samples yield insufficient quantities of particles when using traditional grid making techniques and require the use of solid supports that concentrate samples onto the grid. Recent advances in grid-preparation show that affinity grids are promising tools to selectively concentrate proteins while simultaneously protecting samples from the air-water interface. One such technique utilizes lipid monolayers containing a lipid species with an affinity handle. Some of the first affinity grids used a holey carbon layer coated with nickel nitrilotriacetic acid (Ni-NTA) lipid, which allowed for the binding of proteins bearing the commonly used poly-histidine affinity tag. These studies however used complicated protocols and were conducted before the “resolution revolution” of cryo-EM. Here, we provide a straight-forward preparation method and systematic analysis of Ni-NTA lipid monolayers as a tool for high-resolution single particle cryo-EM. We found that lipid affinity grids concentrate particles away from the air-water interface in thin ice (∼30 nm). We determined a 2.6 Å structure of the human nucleosome, showing this method is amenable to high-resolution structure determination. Furthermore, we determined a 3.1 Å structure of a sub-100 kDa protein demonstrating that this technique is amenable to proteins across biological size ranges. Lipid monolayers are therefore an easily extendable tool for most systems and help alleviate common problems such as low yield, disruption by the air-water interface, and thicker ice.

  PublisherOA PDFDOI

• Structural mechanism of H3K27 demethylation and crosstalk with heterochromatin markers

  Molecular Cell · 2025-07-18 · 2 citations

  articleSenior author
  PublisherDOI

• APC/C-mediated ubiquitylation of extranucleosomal histone complexes lacking canonical degrons

  Nature Communications · 2025-03-15 · 3 citations

  articleOpen accessSenior author

  Non-degradative histone ubiquitylation plays a myriad of well-defined roles in the regulation of gene expression and choreographing DNA damage repair pathways. In contrast, the contributions of degradative histone ubiquitylation on genomic processes has remained elusive. Recently, the APC/C has been shown to ubiquitylate histones to regulate gene expression in pluripotent cells, but the molecular mechanism is unclear. Here we show that despite directly binding to the nucleosome through subunit APC3, the APC/C is unable to ubiquitylate nucleosomal histones. In contrast, extranucleosomal H2A/H2B and H3/H4 complexes are broadly ubiquitylated by the APC/C in an unexpected manner. Using a combination of cryo-electron microscopy (cryo-EM) and biophysical and enzymatic assays, we demonstrate that APC8 and histone tails direct APC/C-mediated polyubiquitylation of core histones in the absence of traditional APC/C substrate degron sequences. Taken together, our work implicates APC/C-nucleosome tethering in the degradation of diverse chromatin-associated proteins and extranucleosomal histones for the regulation of transcription and the cell cycle and for preventing toxicity due to excess histone levels. The APC/C ubiquitylates histones to regulate gene expression in pluripotent cells. Here, the authors pair cryo-EM and biochemical and biophysical assays to show that instead of modifying nucleosome-incorporated histones, the APC/C ubiquitylates extranucleosomal histone complexes through a mechanism that bypasses canonical substrate degrons.

  PublisherOA PDFDOI

• Post-translational modification of H2B C-terminal helix regulates nucleosome interactions and chromatin signaling

  Nucleic Acids Research · 2025-09-04 · 1 citations

  articleOpen accessSenior author

  Histone H2B contains a highly conserved C-terminal (H2B αC) helix that has been implicated in chromatin interactions and dynamics. The H2B αC helix comprising residues 105-125 is positioned adjacent to a major site of nucleosome interactions called the acidic patch. Despite individual structural studies highlighting interactions between chromatin proteins and the H2B αC helix, the general role of the helix in mediating nucleosome recognition has not been explored. Moreover, many post-translational modifications (PTMs) have been identified within the H2B αC helix, but significant gaps exist in our understanding of their regulatory potential. In this study, we employed nucleosome affinity proteomics using a library of nucleosomes with mutations or PTMs of the H2B αC helix to investigate contributions to nucleosome binding. Our work uncovers new spatial patterns of H2B αC helix engagement across the proteome. We also demonstrate that H2B K120 mono-ubiquitylation (H2B K120ub) within the H2B αC helix broadly disrupts nucleosome binding, phenocopying mutation of the acidic patch, while differentially regulating acidic patch-dependent chromatin functions. In contrast, lysine acetylation results in more subtle position-specific changes, highlighting a more general role of H2B αC helix PTMs in tuning acidic patch recognition.

  PublisherDOI

• An Intrinsically Disordered Region of Histone Demethylase Kdm5a Activates Catalysis Through Interactions with the Nucleosomal Acidic Patch and DNA

  SSRN Electronic Journal · 2025-01-01

  preprintOpen access
  PublisherDOI

• Potent and selective SETDB1 covalent negative allosteric modulator reduces methyltransferase activity in cells

  Nature Communications · 2025-02-24 · 6 citations

  articleOpen access

  A promising drug target, SETDB1, is a dual methyl-lysine (Kme) reader and methyltransferase implicated in cancer and neurodegenerative disease progression. To help understand the role of the triple Tudor domain (3TD) of SETDB1, its Kme reader, we first identify a low micromolar potency small molecule ligand, UNC6535, which occupies simultaneously both the TD2 and TD3 reader binding sites. Further optimization leads to the discovery of UNC10013, a covalent 3TD ligand targeting Cys385 of SETDB1. UNC10013 is potent with a kinact/KI of 1.0 × 106 M−1s−1 and demonstrates proteome-wide selectivity. In cells, negative allosteric modulation of SETDB1-mediated Akt methylation occurs after treatment with UNC10013. Therefore, UNC10013 is a potent, selective, and cell-active covalent ligand for the 3TD of SETDB1, demonstrating negative allosteric modulator properties and making it a promising tool to study the biological role of SETDB1 in disease progression. Design of cysteine-targeting analogs of a reversible SETDB1 triple Tudor domain (3TD) ligand, UNC6535, led to UNC10013, a potent covalent ligand with high selectivity. UNC10013 demonstrated allosteric inhibition of SETDB1-mediated Akt methylation in cells, a promising approach to SETDB1 therapeutics.

  PublisherOA PDFDOI

• Nickel-NTA lipid-monolayer affinity grids allow for high-resolution structure determination by cryo-EM

  Journal of Structural Biology · 2025-10-11

  articleOpen access
  PublisherOA PDFDOI

Recent grants

• Molecular Mechanisms of Chromatin Recognition

  NIH · $383k · 2019–2024

• Molecular Mechanisms of Chromatin Recognition

  NIH · $2.5M · 2019–2029

Frequent coauthors

• Aleksandra Skrajna

  University of North Carolina at Chapel Hill

  18 shared

• Tom W. Muir

  Princeton University

  18 shared

• Cathy J. Spangler

  Janelia Research Campus

  11 shared

• Champak Chatterjee

  National Dairy Research Institute

  10 shared

• Robert J. Duronio

  9 shared

• Dennis Goldfarb

  Washington University in St. Louis

  8 shared

• Pengda Liu

  University of North Carolina at Chapel Hill

  8 shared

• Gabrielle R. Budziszewski

  Hauptman-Woodward Medical Research Institute

  8 shared

Labs

• McGinty LabPI

Education

• M.D.

  Weill Cornell Medical College

  2011

• Ph.D.

  Rockefeller University

  2010

• B.S., Biochemistry, Biophysics, and Molecular Biology

  Iowa State University

  2003

Awards & honors

• 2017 Searle Scholar Award
• 2017 Pew-Stewart Scholar for Cancer Research