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Updated: Jun 27, 2025

Author Spotlight: Characterizing DNA Replication of Pathogenic Repeats to Uncover Mechanisms of Replication Fork Stalling and Expansion
05:22

Author Spotlight: Characterizing DNA Replication of Pathogenic Repeats to Uncover Mechanisms of Replication Fork Stalling and Expansion

Published on: September 13, 2024

736

Proteomics Analysis of the Polyomavirus DNA Replication Initiation Complex Reveals Novel Functional Phosphorylated

Rama Dey-Rao1, Shichen Shen2, Jun Qu2

  • 1Department of Microbiology & Immunology, Jacobs School of Medicine & Biomedical Sciences, University at Buffalo, State University of New York at Buffalo, Buffalo, NY 14203, USA.

International Journal of Molecular Sciences
|April 27, 2024
PubMed

Insights

Polyomavirus Large T-antigen (LT) interacts with cellular replication factors, and novel post-translational modifications were identified. This study reveals new insights into viral replication and LT

Area of Science:

  • * Molecular Biology
  • * Virology
  • * Proteomics

Background:

  • * Polyomavirus (PyV) Large T-antigen (LT) is crucial for viral replication and cellular transformation.
  • * LT interacts with cellular replication machinery, including DNA polymerase alpha-primase (Polprim) and replication protein A (RPA).
  • * Post-translational modifications (PTMs) of these factors are known to modulate their activities, but comprehensive proteomic analysis has been limited.

Purpose of the Study:

  • * To comprehensively identify novel PTMs and associated proteins of PyV LT and its interacting replication factors.
  • * To investigate the functional significance of identified PTMs and protein interactions in PyV biology.
  • * To explore the role of transcription factor ETS1 in PyV processes.

Main Methods:

  • * High-resolution liquid chromatography tandem mass spectrometry (LC-MS/MS) was employed for immunoprecipitated factors (IPMS).
  • * Gene Ontology (GO) enrichment analysis was performed for functional annotation.
  • * Coimmunoprecipitation assays were used to validate protein interactions.

Main Results:

  • * IPMS identified 479 novel phosphorylated amino acid residues (PAARs) across the three factors, with one validated functionally.
  • * A large number of novel associated proteins were identified: 374 for LT, 453 for Polprim, and 183 for RPA.
  • * A unique transcription-related network associated with LT was identified, and the transcription factor ETS1 was found to interact with LT, suggesting its involvement in PyV replication.

Conclusions:

  • * The study presents a high-sensitivity proteomic analysis of PyV LT and its interacting replication factors, revealing numerous novel PTMs and protein associations.
  • * The identification of a novel PAAR regulating PyV replication and the association of LT with ETS1 provide valuable insights into PyV biology and pathogenesis.
  • * This work establishes a foundation for further functional studies of identified PTMs and protein interactions in the context of polyomavirus replication and cellular transformation.
Keywords:
DNA replicationDNA virusPolprimRPASV40 LTbioinformaticsnetwork analysisphosphorylationproteomicstranscription factor

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