Dataset Information

Lysine trimethylation in planktonic and pellicle modes of growth in Acinetobacter baumannii

ABSTRACT: An increasing number of studies have shown the importance of post-translational modifications (PTM) in bacterial physiology, especially during biofilm formation. Trimethylation of lysine residues is little studied due to lack of specific enrichment methods for this PTM. However, it may have a significant role in virulence, adaptation or biofilm formation. Acinetobacter baumannii has been described as an important nosocomial pathogen over the past 30 years, frequently implicated in ventilator-associated infections. Many aspects of the biology of A. baumannii remain elusive, in particular the formation of an air-liquid biofilm (pellicle). Here, we investigated K-trimethylation in A. baumannii ATCC 17978 in planktonic and pellicle modes by proteomic analysis, using different analytical approaches. Here, we identified 84 K-trimethylated proteins. Among these proteins, several are localized to the membrane or involved in adhesion, biofilm formation, lipid metabolism and even characterized as virulence or resistance factors. Our results confirmed some K-trimethylated sites previously identified in other bacteria, such as three sites on the 50S ribosomal subunit protein L11 (RplK). But we also identified novel sites such as K1020 on the adhesin Ata autotransporter (Ata), which is a known virulence factor and participates in bacterial adhesion. Overall, this study is the first large-scale proteomic study of trimethylation in A. baumannii and will be an important resource for the scientific community.

INSTRUMENT(S): LTQ Orbitrap Elite

ORGANISM(S): Acinetobacter Baumannii (strain Atcc 17978 / Cip 53.77 / Lmg 1025 / Ncdc Kc755 / 5377)

SUBMITTER: Julie Hardouin

LAB HEAD: Julie Hardouin

PROVIDER: PXD035239 | Pride | 2023-07-20

REPOSITORIES: Pride

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Publications

Lysine Trimethylation in Planktonic and Pellicle Modes of Growth in Acinetobacter baumannii.

Nalpas Nicolas N Kentache Takfarinas T Dé Emmanuelle E Hardouin Julie J

Journal of proteome research 20230525 7

Over the past 30 years, Acinetobacter baumannii has been described as an important nosocomial pathogen due to frequent ventilator-associated infections. Many biological processes of A. baumannii remain elusive, such as the formation of an air-liquid biofilm (pellicle). Several studies demonstrated the importance of post-translational modifications (PTMs) in A. baumannii physiology. Here, we investigated K-trimethylation in A. baumannii ATCC 17978 in planktonic and pel ...[more]

PMID: 37227083

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Project description:Shewanella oneidensis is an important model organism for bioremediation studies because of its diverse respiratory capabilities. In recent years, biofilm development of S. oneidensis has been extensively studied because it is essential to reduce solid metals. As a special form of biofilm, however, pellicles are largely overlooked. The goal of this work was to understand requirements of S. oneidensis pellicle formation and the molecular basis of pellicle formation. We demonstrated that successful pellicle formation and survival was likely to require the threshold level of cell density and higher concentration of oxygen. Proteinase K and EDTA were potent pellicle disrupter. DNA microarray experiments were used to study the gene expression profile of young air–liquid interface pellicle relative to planktonic cells, which indicated that the air–liquid interface pellicle was more metabolically active than the planktonic cells. Most notably, consistently up-regulation of iron or heme uptake and transportation proteins was observed in the S. oneidensis MR-1 pellicle. However, neither the hmuT nor hugA heme transport mutant was defective in pellicle formation. An examination of the influence of several metal cations on the anti-pellicle activity of EDTA showed that Ca (II), Mn(II), Cu(II), and Zn(II) fully protected S. oneidensis MR-1 pellicle against EDTA treatment while additional of iron enabled the initiation of pellicle formation but maturation was significantly impaired. Collectively, iron was less important than other metals with respect to pellicle formation in S. oneidensis. A fresh colony grown overnight on a LB plate was used to inoculate 50 ml LB and incubated in a shaker (200 rpm) to an OD600 of 0.8 at the room temperature. This culture was then diluted 500-fold with fresh LB, resulting in the starting cultures. Aliquots of 30ml starting cultures were transferred to 50-ml Pyrex beakers and allowed to develop pellicles at the room temperature. When a complete but thin (young pellicle) at the interface were formed (about 30h hours), planktonic culture and pellicle were separated and applied to centrifugation at 8000 rpm for 3 min at room temperature. 3 parallel starting cultures were used and 3 samlpes of pellicle cells or planktonic cells were collected at 30h. RNA from the pellicle cells was fluorescently labeled with Cy3, and that from the planktonic was labeled with Cy5.

Dataset Information

Lysine trimethylation in planktonic and pellicle modes of growth in Acinetobacter baumannii

Publications

Lysine Trimethylation in Planktonic and Pellicle Modes of Growth in <i>Acinetobacter baumannii</i>.

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