Dataset Information

A proteomic and functional view of intrabacterial lipid inclusion biogenesis in mycobacteria

ABSTRACT: During infection and granuloma formation, pathogenic mycobacteria accumulate triacylglycerol in the form of Intrabacterial Lipid Inclusions (ILI). These organelles facilitate bacterial evasion from the immune system and serve as a carbon source to sustain bacterial persistence within the infected host. M. abscessus, a rapidly-growing non-tuberculous opportunistic pathogen, was previously shown to generate similar structures throughout their infectious cycle. Herein, it was used as a model organism to identify, at a global level, the ILI-associated proteins (IAP) involved in ILI accumulation. By implementing the APEX2 proximity labeling method using an in vitro model for ILI accumulation, we identified 228 proteins potentially implicated in ILI biosynthesis.

INSTRUMENT(S): Q Exactive Plus

ORGANISM(S): Mycobacterium Tuberculosis Mycobacterium Abscessus

SUBMITTER: AUDEBERT Stephane

LAB HEAD: Stéphane Canaan, Ph. D

PROVIDER: PXD052310 | Pride | 2025-02-27

REPOSITORIES: Pride

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A proteomic and functional view of intrabacterial lipid inclusion biogenesis in mycobacteria.

Dargham Tonia T Aguilera-Correa John Jairo JJ Avellan Romain R Mallick Ivy I Celik Léa L Santucci Pierre P Brasseur Gael G Poncin Isabelle I Point Vanessa V Audebert Stéphane S Camoin Luc L Daher Wassim W Cavalier Jean-François J-F Kremer Laurent L Canaan Stéphane S

mBio 20250225 4

During infection and granuloma formation, pathogenic mycobacteria store triacylglycerol as intrabacterial lipid inclusions (ILIs). This accumulation of nutrients provides a carbon source for bacterial persistence and slows down intracellular metabolism. Mycobacterium abscessus (Mab), a rapidly growing non-tuberculous actinobacterium, produces ILI throughout its infection cycle. Here, Mab was used as a model organism to identify proteins associated with ILI accumulation on a ...[more]

PMID: 39998225

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Project description:To study the molecular mediators of naturally rewarding effects of palatable food we used a model of palatable âsnackingâ (Ulrich-Lai et al., 2007) in which rats are given chronic, brief access to a limited amount of sucrose solution (30%). Single housed, male Long-Evans rats (250g) (n=12 per group) from Harlan Labs (Indianapolis, IN) received normal rat chow (Harlan Teklad) and water ad libitum for the duration of the experiment. After a one-week period of acclimation, rats were randomly assigned to drink treatment groups of either 30% sucrose solution or water. Rats received a 14-day regimen of twice daily (9:30 and 15:30) brief (maximum of 30 minutes) limited (up to 4 mL) access of their assigned drink solution. Drink solutions were delivered via a graduated sipper placed onto the cage top in addition to the existing water bottle and sippers were immediately removed when the animal had consumed 4mL or after the 30-minute access period, whichever occurred first. Drink intake, food intake, and body weight were monitored throughout the experiment to verify that the rats learned to drink sucrose, that they adjusted chow intake for calories consumed from sucrose (~10%), and that there was no effect on body weight gain as is normally seen with this model (Ulrich-Lai et al., 2007). Drink treatment terminated on day 14 and at 8:00 on the morning of day 15, the rats were sacrificed by rapid decapitation. BLA tissue was dissected, RNA extracted, and gene expression changes between water and sucrose groups were accessed by microarray. We used microarray to assess the changes in gene expression in the BLA of rats that have had a history of sucrose snacks (n=12) vs control (n=12). Rats received access (up to 4 mL) to extra bottles containing water (control) or 30% sucrose twice daily for 14 days. Rats had ab lib access to chow and water for the duration of the experiment. Rats were sacrificed the morning of day 15 and the BLA was removed for microarray analysis. Functional cluster analysis was performed on the list of 145 significantly upregulated genes using Ingenuity Pathway Analysis (Ingenuity Systems, Redwood City, CA). See supplementary file at the foot of this record.

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