Dataset Information

Proteomic response of Deinococcus radiodurans to short-term real microgravity during parabolic flight

ABSTRACT: As plans for future space exploration are becoming more ambitious, a better understanding of all factors affecting humans, plants, and microorganisms in space is necessary. Microgravity is an important variable in outer space and understanding the short- and long- term effects of microgravity on cellular processes will be important to minimize its negative effects on the physiology of any organism. Gravitational force has had an important role in the development of life on Earth, and short- and long-term changes in perceived gravitational force can induce notable changes within cells. Deinococcus radiodurans is the gram-positive bacterium that is best known for its extreme resistance to UV-C and gamma radiation, oxidation stress and desiccation, which has led to increased interest in this species in the context of astrobiology. The present study aimed to elucidate the short-term proteomic response of this species to real microgravity during parabolic flight. Overnight cultures were subjected to microgravity during a single parabola, and metabolic activity quenched using methanol. Proteins were extracted and subsequently measured using HPLC ESI MS/MS. Results indicated multiple affected processes in the cell envelope of D. radiodurans, such as increased peptidoglycan synthesis and altered S-layer activities. Energy metabolism upregulation and increased activity of DNA repair pathways could indicate increased endogenous ROS production that contributes to the stress response. The present study shows that the D. radiodurans proteome reacts to real microgravity within seconds. Differential expression patterns in response to microgravity show similarities to previously reported stress responses, thus the present results could be used as basis for future research aiming to better understand the complex regulatory processes underpinning stress management in D. radiodurans.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Deinococcus Radiodurans R1

SUBMITTER: Karlis Moors

LAB HEAD: Dr Tetyana Milojevic

PROVIDER: PXD027236 | Pride | 2022-02-17

REPOSITORIES: Pride

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Proteomic Response of <i>Deinococcus radiodurans</i> to Short-Term Real Microgravity during Parabolic Flight Reveals Altered Abundance of Proteins Involved in Stress Response and Cell Envelope Functions.

Moors Karlis Arturs KA Ott Emanuel E Weckwerth Wolfram W Milojevic Tetyana T

Life (Basel, Switzerland) 20211224 1

Rapidly evolving space exploration makes understanding the short- and long- term effects of microgravity on humans, plants, and microorganisms an important task. The ubiquitous presence of the gravitational force has had an influence on the development of all living entities on Earth, and short- and long-term changes in perceived gravitational force can induce notable changes within cells. <i>Deinococcus radiodurans</i> is the Gram-positive bacterium that is best known for its extreme resistance ...[more]

PMID: 35054415

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Project description:Abstract Background The extraordinarily resistant bacterium Deinococcus radiodurans withstands harsh environmental conditions present in outer space. Deinococcus radiodurans was exposed for one year outside the International Space Station within Tanpopo orbital mission to investigate microbial survival and space travel. In addition, a ground-based simulation experiment with conditions, mirroring those from Low Earth orbit, was performed. Methods We monitored Deinococcus radiodurans cells during early stage of recovery after Low Earth orbit exposure using electron microscopy tools. Furthermore, proteomic, transcriptomic and metabolomic analyses were performed to identify molecular mechanisms responsible for the survival of Deinococcus radiodurans in Low Earth orbit. Results D. radiodurans cells exposed to low Earth orbit conditions do not exhibit any morphological damage. However, an accumulation of numerous outer-membrane associated vesicles was observed. On levels of proteins and transcripts, a multifaceted response was detected to alleviate cell stress. The UvrABC endonuclease excision repair mechanism was triggered to cope with DNA damage. Defense against reactive oxygen species is mirrored by the increased abundance of catalases and is accompanied by the increased abundance of putrescine which works as scavenging molecule. In addition, several proteins and mRNAs, responsible for regulatory and transporting functions showed increased abundances. The decrease in primary metabolites indicate alternations in the energy status, which is needed to repair damaged molecules. Conclusion Low Earth orbit induced molecular rearrangements trigger multiple components of metabolic stress response and regulatory networks in exposed microbial cells. Presented results show that the non-sporulating bacterium Deinococcus radiodurans survived long-term Low Earth orbit exposure if wavelength below 200 nm are not present, which mirrors the UV spectrum of Mars, where CO2 effectively provides a shield below 190 nm. These results should be considered in the context of planetary protection concerns and the development of new sterilization techniques for future space missions.

Project description:Characterization of bacterial behavior in the microgravity environment of spaceflight is of importance towards risk assessment and prevention of infectious disease during long-term missions. Further, this research field unveils new insights into connections between low fluid-shear regions encountered by pathogens during their natural infection process in vivo, and bacterial virulence. This study is the first to characterize the global transcriptomic and proteomic response of an opportunistic pathogen that is actually found in the space habitat, Pseudomonas aeruginosa. Overall, P. aeruginosa responded to spaceflight conditions through differential regulation of 167 genes and 28 proteins, with Hfq identified as a global transcriptional regulator in the response to this environment. Since Hfq was also induced in spaceflight-grown Salmonella typhimurium, Hfq represents the first spaceflight-induced regulator across the bacterial species border. The major P. aeruginosa virulence-related genes induced in spaceflight conditions were the lecA and lecB lectins and the rhamnosyltransferase (rhlA), involved in the production of rhamnolipids. The transcriptional response of spaceflight-grown P. aeruginosa was compared with our previous data of this organism grown in microgravity-analogue conditions using the rotating wall vessel (RWV) bioreactor technology. Interesting similarities were observed, among others with regard to Hfq regulation and oxygen utilization. While LSMMG-grown P. aeruginosa mainly induced genes involved in microaerophilic metabolism, P. aeruginosa cultured in spaceflight adopted an anaerobic mode of growth, in which denitrification was presumably most prominent. Differences in hardware between spaceflight and LSMMG experiments, in combination with more pronounced low fluid shear and mixing in spaceflight when compared to LSMMG conditions, were hypothesized to be at the origin of these observations. Collectively, our data suggest that spaceflight conditions could induce the transition of P. aeruginosa from an opportunistic organism to potential pathogen, results that are of importance for infectious disease risk assessment and prevention, both during spaceflight missions and in the clinic. This study describes the transcriptional response of P. aeruginosa PAO1 to low-Earth orbit environmental conditions. Our aim was to assess whether the microgravity environment of spaceflight could induce virulence traits in P. aeruginosa. To this end, P. aeruginosa cultures were grown in space, and the expression profile was compared with ground control samples (both in biological triplicate). Two RWV samples also examined (did not re-analyze them, only compared the outputs).

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