Systematic Analysis of the Lysine Crotonylome and Multiple Posttranslational Modification Analysis (Acetylation, Succinylation, and Crotonylation) in Candida albicans

Abstract

Candida albicans is an opportunistic pathogen that causes lethal fungal infections in immunocompromised patients. Lysine crotonylation is a newly discovered PTM (posttranslational modification) epigenetic type that may play a critical role in regulating gene expression. In this study, we used an antibody-enrichment approach along with LC-MS/MS to carry out a quantitative crotonylome analysis in C. albicans. We found a total of 5,242 crotonylation sites and 1,584 crotonylated proteins among 9,038 proteins in this organism. Of these crotonylated proteins, a few unique crotonylated motifs are noted such as D and E in positions +1, +2, or +3 or K and R in positions +5 or +6, while A, E, F, G, P, W, and Y are in the −1 position or A, K, and R are found in positions −5, −6, −7, or −8. Functional analysis has shown that a majority of the crotonylated proteins are related to biosynthetic events and carbon metabolism. When combined with previously collected data on acetylation and succinylation, PPI (protein-protein interaction network) analysis reveals that proteins with functions in ribosomal biogenesis, oxidative phosphorylation, nucleus activity, and proteasome formation are heavily modified by these three PTM types. To the best of our knowledge, this is the first crotonylome study carried out in C. albicans and is an important step to a better understanding of the biological and pathogenic impact of PTM in C. albicans. IMPORTANCE C. albicans is a kind of pathogen of fungal infections that is found worldwide. Lysine crotonylation of proteins as a recently discovered PTM (posttranslational modification) may have a critical role in regulating cells. We first carried out large-scale analysis of crotonylated proteome and multiple PTM analysis (acetylation, succinylation, and crotonylation), then drew a diagram to show multiple PTM sites on histones in C. albicans of our study. This study about crotonylome in human pathogenic fungi is a milestone that first and deeply investigates the functional analysis of crotonylated proteins in C. albicans, which marks an important start for further research.

Dynamics and Functional Interplay of Nonhistone Lysine Crotonylome and Ubiquitylome in Vascular Smooth Muscle Cell Phenotypic Remodeling

“…In the past research, there are articles summarizing the cross talk of multiple PTMs in Candida albicans ( 51 ), developing rice seeds ( 52 ), yeast sporulation, and mouse spermatogenesis ( 53 ). In VSMC, little is known about the cross talk of multi-PTMs.…”

Dynamics and Functional Interplay of Non-histone Lysine Crotonylome and Ubiquitylome in Vascular Smooth Muscle Cell Phenotypic Remodeling

“…In the past research, there are articles summarizing the crosstalk of multiple post-translational modi cations in Candida albicans [52], developing rice seeds [53], yeast sporulation and mouse spermatogenesis [54]. In VSMC, little is known about the crosstalk of multi-post-translational modi cations.…”

Pathogen-Host Interaction Repertoire at Proteome and Posttranslational Modification Levels During Fungal Infections

“…Third, interactions of proteomes and regulation mechanisms among PTMs are poorly understood. In addition, although proteomics together with other omics can serve as comprehensive displays of cellular transcriptional levels, unfortunately, most multiomic studies are presented without simultaneous analyses and functional experiments ( Zamith-Miranda et al., 2019 ; Zhou et al., 2021 ). This lack of supporting studies may be due to constraints with database integration and interconnectivity of omics data ( Song M. et al., 2020 ).…”

“…Over past decades, application of MS-based proteomics has expanded rapidly, especially in studies of proteomes and posttranslational modifications (PTMs), such as acetylation, phosphorylation, succinylation, and crotonylation ( Figure 1A ) ( Aggarwal et al., 2021 ). Application in studies of microbiological pathogenesis and interactions between pathogens and hosts has led to the discovery of many novel mechanisms of host–fungus interactions ( Toor et al., 2018 ; Khan et al., 2019 ; Li Y. et al., 2019 ; Zamith-Miranda et al., 2019 ; Bruno et al., 2020 ; Machata et al., 2020 ; Thak et al., 2020 ; Zhou et al., 2021 ). Establishing connections between proteomic profiles and fungal infection processes is critical in characterizing disease pathophysiology, developing candidate therapies, and predicting clinical outcomes.…”