Identification of lysine acetylation sites on MERS-CoV replicase pp1ab

Lin Zhu, Sin Yee Fung, Guangshan Xie, Lok Yin Roy Wong, Dong Yan Jin, Zongwei CAI*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

3 Citations (Scopus)

Abstract

MERS is a life-threatening disease and MERS-CoV has the potential to cause the next pandemic. Protein acetylation is known to play a crucial role in host response to viral infection. Acetylation of viral proteins encoded by other RNA viruses have been reported to affect viral replication. It is therefore of interest to see whether MERS-CoV proteins are also acetylated. Viral proteins obtained from infected cells were trypsin-digested into peptides. Acetylated peptides were enriched by immunoprecipitation and subject to nano-LC-Orbitrap analysis. Bioinformatic analysis was performed to assess the conservation level of identified acetylation sites and to predict the upstream regulatory factors. A total of 12 acetylation sites were identified from 7 peptides, which all belong to the replicase polyprotein pp1ab. All identified acetylation sites were found to be highly conserved across MERS-CoV sequences in NCBI database. Upstream factors, including deacetylases of the SIRT1 and HDAC families as well as acetyltransferases of the TIP60 family, were predicted to be responsible for regulating the acetylation events identified. Western blot confirms that acetylation events indeed occur on pp1ab protein by expressing NSP4 in HEK293 cells. Acetylation events on MERS-CoV viral protein pp1ab were identified for the first time, which indicate that MERS-CoV might use the host acetylation machinery to regulate its enzyme activity and to achieve optimal replication. Upstream factors were predicted, which might facilitate further analysis of the regulatory mechanism of MERS-CoV replication.

Original languageEnglish
JournalMolecular and Cellular Proteomics
Volume19
Issue number8
DOIs
Publication statusPublished - Aug 2020

Scopus Subject Areas

  • Analytical Chemistry
  • Biochemistry
  • Molecular Biology

User-Defined Keywords

  • Acetylation
  • Bioinformatics
  • MERS-CoV
  • PTM proteomics
  • Virus-host interaction

Fingerprint

Dive into the research topics of 'Identification of lysine acetylation sites on MERS-CoV replicase pp1ab'. Together they form a unique fingerprint.

Cite this