Elucidating Proteoform Dynamics Underlying the Senescence Associated Secretory Phenotype

Peter F. Doubleday, Luca Fornelli, Neil L. Kelleher*

*Corresponding author for this work

Research output: Contribution to journalArticle

Abstract

Primary diploid cells exit the cell cycle in response to exogenous stress or oncogene activation through a process known as cellular senescence. This cell-autonomous tumor-suppressive mechanism is also a major mechanism operative in organismal aging. To date, temporal aspects of senescence remain understudied. Therefore, we use quantitative proteomics to investigate changes following forced HRASG12V expression and induction of senescence across 1 week in normal diploid fibroblasts. We demonstrate that global intracellular proteomic changes correlate with the emergence of the senescence-associated secretory phenotype and the switch to robust cell cycle exit. The senescence secretome reinforces cell cycle exit, yet is largely detrimental to tissue homeostasis. Previous studies of secretomes rely on ELISA, bottom-up proteomics or RNA-seq. To date, no study to date has examined the proteoform complexity of secretomes to elucidate isoform-specific, post-translational modifications or regulated cleavage of signal peptides. Therefore, we use a quantitative top-down proteomics approach to define the molecular complexity of secreted proteins <30 kDa. We identify multiple forms of immune regulators with known activities and affinities such as distinct forms of interleukin-8, as well as GROα and HMGA1, and temporally resolve secreted proteoform dynamics. Together, our work demonstrates the complexity of the secretome past individual protein accessions and provides motivation for further proteoform-resolved measurements of the secretome.

Original languageEnglish (US)
Pages (from-to)938-948
Number of pages11
JournalJournal of Proteome Research
Volume19
Issue number2
DOIs
StatePublished - Feb 7 2020

Keywords

  • proteoform
  • quantitative proteomics
  • secretome
  • senescence
  • top-down proteomics

ASJC Scopus subject areas

  • Biochemistry
  • Chemistry(all)

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