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Identification and characterisation of sPEPs in Cryptococcus neoformans.

Paige E Erpf1, Sheena M H Chua1, Toan K Phung1

  • 1The University of Queensland, School of Chemistry & Molecular Biosciences, St Lucia, Queensland 4072, Australia; The University of Queensland, Australian Infectious Diseases Research Centre, School of Chemistry & Molecular Biosciences, St Lucia, Queensland 4072, Australia.

Fungal Genetics and Biology : FG & B
|March 27, 2022
PubMed
Summary

Researchers identified novel short open reading frame-encoded peptides (sPEPs) in Cryptococcus neoformans using a new enrichment and proteogenomic approach. One discovered sPEP, Npb1, functions similarly to bacterial trans-translation proteins.

Keywords:
Cryptococcus neoformansFungiGeneticsPathogenProteogenomicsProteomicsSmall peptidessPEPs

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Area of Science:

  • Microbiology
  • Molecular Biology
  • Proteomics

Background:

  • Short open reading frame-encoded peptides (sPEPs) are increasingly recognized across species.
  • Identification and characterization of sPEPs in Cryptococcus neoformans have been hindered by a lack of standardized methods.

Purpose of the Study:

  • To develop and implement an enrichment and proteogenomic strategy for sPEP detection in C. neoformans.
  • To identify and validate novel sORFs and their encoded sPEPs within the C. neoformans genome.

Main Methods:

  • Developed a novel enrichment process for sPEP detection in C. neoformans protein samples.
  • Utilized proteogenomics to validate predicted and hypothetical sORFs.
  • Generated specific point mutations and a full deletion allele for functional characterization of a novel sPEP (Npb1).

Main Results:

  • Successfully enabled sPEP detection in the polysaccharide-encapsulated yeast C. neoformans.
  • Discovered novel sORFs in 5' and 3' UTRs of known transcripts and within non-coding RNAs.
  • Characterized a novel sPEP, Npb1, demonstrating functional similarity to bacterial SmpB.

Conclusions:

  • The developed proteogenomic approach is effective for sPEP discovery in C. neoformans.
  • Identified novel sPEPs, including Npb1, expanding the known functional landscape of this fungal pathogen.
  • Npb1 represents a novel sPEP with a conserved function related to bacterial trans-translation.