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  1. Article: Analyses of cell wall synthesis in

    Shrestha, Shailab / Taib, Najwa / Gribaldo, Simonetta / Shen, Aimee

    bioRxiv : the preprint server for biology

    2023  

    Abstract: Current models of bacterial cell division assume that the core synthases of the multiprotein divisome complex, FtsW-FtsI, are the primary drivers of septal peptidoglycan (PG) synthesis. These enzymes are typically encoded in the highly conserved division ...

    Abstract Current models of bacterial cell division assume that the core synthases of the multiprotein divisome complex, FtsW-FtsI, are the primary drivers of septal peptidoglycan (PG) synthesis. These enzymes are typically encoded in the highly conserved division and cell wall (
    Language English
    Publishing date 2023-08-08
    Publishing country United States
    Document type Preprint
    DOI 10.1101/2023.08.06.552200
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  2. Article ; Online: Diversification of division mechanisms in endospore-forming bacteria revealed by analyses of peptidoglycan synthesis in Clostridioides difficile.

    Shrestha, Shailab / Taib, Najwa / Gribaldo, Simonetta / Shen, Aimee

    Nature communications

    2023  Volume 14, Issue 1, Page(s) 7975

    Abstract: The bacterial enzymes FtsW and FtsI, encoded in the highly conserved dcw gene cluster, are considered to be universally essential for the synthesis of septal peptidoglycan (PG) during cell division. Here, we show that the pathogen Clostridioides ... ...

    Abstract The bacterial enzymes FtsW and FtsI, encoded in the highly conserved dcw gene cluster, are considered to be universally essential for the synthesis of septal peptidoglycan (PG) during cell division. Here, we show that the pathogen Clostridioides difficile lacks a canonical FtsW/FtsI pair, and its dcw-encoded PG synthases have undergone a specialization to fulfill sporulation-specific roles, including synthesizing septal PG during the sporulation-specific mode of cell division. Although these enzymes are directly regulated by canonical divisome components during this process, dcw-encoded PG synthases and their divisome regulators are dispensable for cell division during normal growth. Instead, C. difficile uses a bifunctional class A penicillin-binding protein as the core divisome PG synthase, revealing a previously unreported role for this class of enzymes. Our findings support that the emergence of endosporulation in the Firmicutes phylum facilitated the functional repurposing of cell division factors. Moreover, they indicate that C. difficile, and likely other clostridia, assemble a distinct divisome that therefore may represent a unique target for therapeutic interventions.
    MeSH term(s) Bacterial Proteins/genetics ; Bacterial Proteins/metabolism ; Clostridioides difficile/genetics ; Clostridioides difficile/metabolism ; Peptidoglycan/metabolism ; Membrane Proteins/metabolism ; Endospore-Forming Bacteria/metabolism
    Chemical Substances Bacterial Proteins ; Peptidoglycan ; Membrane Proteins
    Language English
    Publishing date 2023-12-02
    Publishing country England
    Document type Journal Article
    ZDB-ID 2553671-0
    ISSN 2041-1723 ; 2041-1723
    ISSN (online) 2041-1723
    ISSN 2041-1723
    DOI 10.1038/s41467-023-43595-3
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  3. Article ; Online: The influenza A virus host shutoff factor PA-X is rapidly turned over in a strain-specific manner.

    Levene, Rachel Emily / Shrestha, Shailab D / Gaglia, Marta Maria

    Journal of virology

    2021  Volume 95, Issue 8

    Abstract: The influenza A endoribonuclease PA-X regulates virulence and transmission of the virus by reducing host gene expression and thus regulating immune responses to influenza A virus. Despite this key function in viral biology, the levels of PA-X protein ... ...

    Abstract The influenza A endoribonuclease PA-X regulates virulence and transmission of the virus by reducing host gene expression and thus regulating immune responses to influenza A virus. Despite this key function in viral biology, the levels of PA-X protein remain markedly low during infection, and previous results suggest that these low levels are not solely the result of regulation of the level of translation and RNA stability. How PA-X is regulated post-translationally remains unknown. We now report that the PA-X protein is rapidly turned over. PA-X from multiple viral strains are short-lived, although the half-life of PA-X ranges from ∼30 minutes to ∼3.5 hours depending on the strain. Moreover, sequences in the variable PA-X C-terminal domain are primarily responsible for regulating PA-X half-life, although the N-terminal domain also accounts for some differences among strains. Interestingly, we find that the PA-X from the 2009 pandemic H1N1 strain has a longer half-life compared to the other variants we tested. This PA-X isoform has been reported to have a higher host shutoff activity, suggesting a role for protein turnover in regulating PA-X activity. Collectively, this study reveals a novel regulatory mechanism of PA-X protein levels that may impact host shutoff activity during influenza A virus infection.
    Language English
    Publishing date 2021-01-27
    Publishing country United States
    Document type Journal Article
    ZDB-ID 80174-4
    ISSN 1098-5514 ; 0022-538X
    ISSN (online) 1098-5514
    ISSN 0022-538X
    DOI 10.1128/JVI.02312-20
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  4. Article ; Online: Single-spore germination analyses reveal that calcium released during

    Ribis, John W / Melo, Luana / Shrestha, Shailab / Giacalone, David / Rodriguez, Enrique E / Shen, Aimee / Rohlfing, Amy

    mSphere

    2023  Volume 8, Issue 4, Page(s) e0000523

    Abstract: Clostridioides ... ...

    Abstract Clostridioides difficile
    MeSH term(s) Calcium/metabolism ; Clostridioides/metabolism ; Clostridioides difficile/physiology ; Spores, Bacterial/physiology ; Bacterial Proteins/genetics ; Bacterial Proteins/metabolism ; Amino Acids/metabolism ; Bile Acids and Salts/pharmacology ; Bile Acids and Salts/metabolism
    Chemical Substances Calcium (SY7Q814VUP) ; Bacterial Proteins ; Amino Acids ; Bile Acids and Salts
    Language English
    Publishing date 2023-06-20
    Publishing country United States
    Document type Journal Article
    ISSN 2379-5042
    ISSN (online) 2379-5042
    DOI 10.1128/msphere.00005-23
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  5. Article ; Online: Development of a Dual-Fluorescent-Reporter System in Clostridioides difficile Reveals a Division of Labor between Virulence and Transmission Gene Expression.

    Donnelly, M Lauren / Shrestha, Shailab / Ribis, John W / Kuhn, Pola / Krasilnikov, Maria / Alves Feliciano, Carolina / Shen, Aimee

    mSphere

    2022  Volume 7, Issue 3, Page(s) e0013222

    Abstract: The bacterial pathogen Clostridioides difficile causes gastroenteritis by producing toxins and transmits disease by making resistant spores. Toxin and spore production are energy-expensive processes that are regulated by multiple transcription factors in ...

    Abstract The bacterial pathogen Clostridioides difficile causes gastroenteritis by producing toxins and transmits disease by making resistant spores. Toxin and spore production are energy-expensive processes that are regulated by multiple transcription factors in response to many environmental inputs. While toxin and sporulation genes are both induced in only a subset of C. difficile cells, the relationship between these two subpopulations remains unclear. To address whether C. difficile coordinates the generation of these subpopulations, we developed a dual-transcriptional-reporter system that allows toxin and sporulation gene expression to be simultaneously visualized at the single-cell level using chromosomally encoded mScarlet and mNeonGreen fluorescent transcriptional reporters. We then adapted an automated image analysis pipeline to quantify toxin and sporulation gene expression in thousands of individual cells under different medium conditions and in different genetic backgrounds. These analyses revealed that toxin and sporulation gene expression rarely overlap during growth on agar plates, whereas broth culture increases this overlap. Our results suggest that certain growth conditions promote a "division of labor" between transmission and virulence gene expression, highlighting how environmental inputs influence these subpopulations. Our data further suggest that the RstA transcriptional regulator skews the population to activate sporulation genes rather than toxin genes. Given that recent work has revealed population-wide heterogeneity for numerous cellular processes in C. difficile, we anticipate that our dual-reporter system will be broadly useful for determining the overlap between these subpopulations.
    MeSH term(s) Agar ; Bacterial Proteins/genetics ; Bacterial Proteins/metabolism ; Bacterial Toxins/genetics ; Bacterial Toxins/metabolism ; Clostridioides ; Clostridioides difficile/genetics ; Gene Expression ; Gene Expression Regulation, Bacterial ; Spores, Bacterial ; Virulence
    Chemical Substances Bacterial Proteins ; Bacterial Toxins ; Agar (9002-18-0)
    Language English
    Publishing date 2022-05-31
    Publishing country United States
    Document type Journal Article
    ISSN 2379-5042
    ISSN (online) 2379-5042
    DOI 10.1128/msphere.00132-22
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  6. Article: Draft Genome Sequences of 10 Environmental

    Shrestha, Shailab D / Guttman, David S / Perron, Gabriel G

    Genome announcements

    2017  Volume 5, Issue 34

    Abstract: Pseudomonas ... ...

    Abstract Pseudomonas aeruginosa
    Language English
    Publishing date 2017-08-24
    Publishing country United States
    Document type Journal Article
    ZDB-ID 2704277-7
    ISSN 2169-8287
    ISSN 2169-8287
    DOI 10.1128/genomeA.00804-17
    Database MEDical Literature Analysis and Retrieval System OnLINE

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