# Administrative supplement to investigate the localization of DNA alkylation damage repair complexes via ubiquitin signaling

> **NIH NIH R15** · MOUNT ST. MARY'S UNIVERSITY · 2024 · $52,180

## Abstract

Project Summary/Abstract
Over the past 20 years, the protein ASCC2 has been shown to contribute to the localization of numerous
multiprotein complexes involved in pathways such as DNA alkylation damage repair and the stalled ribosome
response. A common feature of several of these pathways is that they assemble K63-linked polyubiquitin chains
which are recognized by ASCC2 to recruit the appropriate multiprotein complexes. ASCC2 utilizes a CUE domain
to bind the K63-linked polyubiquitin chains, however, CUE domains are typically promiscuous ubiquitin binders,
and it is unclear how ASCC2 specifically recognizes K63-linked polyubiquitin chains among the myriad types of
polyubiquitin chains present in the cell. The overarching goal of the proposed experiments is to elucidate the
structural basis for ASCC2’s specificity for binding K63-linked polyubiquitin chains with the following specific
aims: 1) identify the intermolecular interactions that mediate binding between ASCC2 and K63-linked
polyubiquitin chains using nuclear magnetic resonance (NMR) spectroscopy, 2) use integrative modeling to
create a representation of the interaction between ASCC2 and K63-linked polyubiquitin chains that agrees with
our experimentally determined NMR, mutagenesis, and small angle X-ray scattering data, and 3) quantify the
functional significance of the predicted interactions between ASCC2 and K63-linked polyubiquitin chains using
in vitro and cell-based assays. These studies are part of a broader effort to better understand the biological
pathways that depend on ASCC2’s ubiquitin-binding abilities and to diagnose diseases associated with the
malfunction of these pathways. For example, in the DNA alkylation damage repair pathway, mutations that inhibit
the formation of the K63-linked polyubiquitin chains that recruit ASCC2, and mutations that inhibit the interaction
between ASCC2 and other members of the ALKBH3-ASCC DNA repair complex, have recently been associated
with genetic diseases due to research in this area. Additionally, the proposed studies will provide valuable
research opportunities for students at Mount St. Mary’s University (MSMU). Collaborations with scientists at
Johns Hopkins University, Washington University in St. Louis, and the SIBYLS beamline at the Lawrence
Berkeley National Laboratory will allow the MSMU undergraduate researchers access to scientific
instrumentation that is not available on their home campus. Overall, the proposed studies will further our
understanding of how ASCC2 targets multiprotein complexes to sites marked by K63-linked polyubiquitin chains
while greatly enhancing the research opportunities available for students at MSMU.

## Key facts

- **NIH application ID:** 11035321
- **Project number:** 3R15GM140410-02S1
- **Recipient organization:** MOUNT ST. MARY'S UNIVERSITY
- **Principal Investigator:** Patrick Lombardi
- **Activity code:** R15 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $52,180
- **Award type:** 3
- **Project period:** 2024-02-15 → 2024-12-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/11035321

## Citation

> US National Institutes of Health, RePORTER application 11035321, Administrative supplement to investigate the localization of DNA alkylation damage repair complexes via ubiquitin signaling (3R15GM140410-02S1). Retrieved via AI Analytics 2026-05-25 from https://api.ai-analytics.org/grant/nih/11035321. Licensed CC0.

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