The Novel Membrane-Associated Auxiliary Factors AuxA and AuxB Modulate β-lactam Resistance in MRSA by stabilizing Lipoteichoic Acids

Mikkelsen, Kasper; Sirisarn, Wanchat; Alharbi, Ohood; Alharbi, Mohanned Talal; Liu, Huayong; Nøhr-Meldgaard, Katrine; Mayer, Katharina; Vestergaard, Martin; Gallagher, Laura A.; Derrick, Jeremy P.; McBain, Andrew J.; Biboy, Jacob; Vollmer, Waldemar; O’Gara, James P.; Grunert, Tom; Ingmer, Hanne; Xia, Guoqing

doi:10.1016/j.ijantimicag.2021.106283

Cited by 27 publications

(34 citation statements)

References 39 publications

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“…The same is also the case for a diversity of factors contributing to cell division and cell wall biogenesis, including FtsZ, FtsA, PBP4 and most proteins involved in the PG synthesis pathway (55). The secretion-associated proteins SecDF and the chaperones PrsA and HtrA1 have also been tightly linked to β-lactam sensitivity (57, 58), and recently, inactivation of the autolytic cell wall amidase Sle1 (21), or the membrane proteins AuxA and AuxB (44), were shown to result in increased sensitivity to β-lactams. Mechanisms of re-sensitization in these cases vary and may be a result of weakened cell wall, as well as inactivation or mis-localization of PG synthesis, including the key resistance determinant PBP2A.…”

Section: Discussionmentioning

confidence: 99%

“…S7A). We also performed Fourier transform infrared spectroscopy (FTIR), which has been used before to detect differences in the composition of WTA due to variable glycosylation patterns (Grunert et al, 2018;Mikkelsen et al, 2021). As expected, changes in the FTIR spectra were evident in the polysaccharide region (1200 cm -1 -800 cm -1 ) for the TarO down strain compared to the control, with the most significant differences recorded for the peaks at 1076 cm -1 , 1048 cm -1 , 1033 cm -1 and 1000 -970 cm -1 , representing α-and β-glycosidic bonds in WTA (Grunert et al, 2018).…”

Section: Smda Has No Major Effects On the Ta Biosynthetic Pathwaysmentioning

confidence: 99%

“…Same procedure as in (Mikkelsen et al, 2021) was carried out for possible detection of LTA released to the medium. The supernatants, after harvesting cells during sample preparations, were kept.…”

Section: Detection Of Lipoteichoic Acid (Lta) By Western Blottingmentioning

confidence: 99%

“…The membrane was again washed three times and LTA bands were visualized by using SuperSignal TM West Pico PLUS Chemiluminescent substrate (Thermo Fisher Scientific) in an Azure Imager c400 (Azure Biosystems). Same procedure as in (44) was carried out for possible detection of LTA released to the medium. The supernatants, after harvesting cells during sample preparations, were kept.…”

Section: Detection Of Lipoteichoic Acid (Lta) By Western Blottingmentioning

confidence: 99%

“…, and recently, inactivation of the autolytic cell wall amidase Sle1(Thalsø-Madsen et al, 2019), or the membrane proteins AuxA and AuxB(Mikkelsen et al, 2021), were shown to result in increased sensitivity to β-lactams. Mechanisms of re-sensitization in these cases vary and may be a result of weakened cell wall, as well as inactivation or mis-localization of PG synthesis, including the key resistance determinant PBP2A.…”

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SmdA is a novel cell morphology determinant in Staphylococcus aureus

Myrbråten

Stamsås

Chan

et al. 2021

Preprint

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Cell division and cell wall synthesis in staphylococci need to be precisely coordinated and controlled to allow the cell to multiply while maintaining their nearly spherical shape. The mechanisms ensuring correct placement of the division plane and synthesis of new cell wall have been studied intensively, however, hitherto unknown factors and proteins are likely to play key roles in this complex interplay. We here identified and investigated a protein with major influence on cell morphology in Staphylococcus aureus. The protein, named SmdA (for staphylococcal morphology determinant A), is a membrane-protein with septum-enriched localization. By CRISPRi knockdown and overexpression combined with different microscopy techniques, we demonstrate that proper levels of SmdA is necessary for cell division, including septum formation and cell splitting. We also identified conserved residues in SmdA that are critical for its functionality. Pulldown- and bacterial two-hybrid interaction experiments showed that SmdA interacts with several known cell division- and cell wall synthesis proteins, including penicillin binding proteins (PBPs) and EzrA. Notably, SmdA also affects susceptibility to cell wall targeting antibiotics, particularly in methicillin-resistant S. aureus (MRSA). Together, our results show that S. aureus is dependent on balanced amounts of membrane-attached SmdA in order to carry out proper cell division.

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Section: Discussionmentioning

confidence: 99%

Section: Smda Has No Major Effects On the Ta Biosynthetic Pathwaysmentioning

confidence: 99%

Section: Detection Of Lipoteichoic Acid (Lta) By Western Blottingmentioning

confidence: 99%

Section: Detection Of Lipoteichoic Acid (Lta) By Western Blottingmentioning

confidence: 99%

mentioning

confidence: 99%

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SmdA is a novel cell morphology determinant in Staphylococcus aureus

Myrbråten

Stamsås

Chan

et al. 2021

Preprint

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Controlling biofilm and virulence properties of Gram-positive bacteria by targeting wall teichoic acid and lipoteichoic acid

Jeong

Khan

Tabassum

et al. 2023

International Journal of Antimicrobial Agents

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Rapid Inhibitor Discovery by Exploiting Synthetic Lethality

et al. 2022

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Synthetic lethality occurs when inactivation of two genes is lethal but inactivation of either single gene is not. This phenomenon provides an opportunity for efficient compound discovery. Using differential growth screens, one can identify biologically active compounds that selectively inhibit proteins within the synthetic lethal network of any inactivated gene. Here, based purely on synthetic lethalities, we identified two compounds as the only possible inhibitors of Staphylococcus aureus lipoteichoic acid (LTA) biosynthesis from a screen of ∼230,000 compounds. Both compounds proved to inhibit the glycosyltransferase UgtP, which assembles the LTA glycolipid anchor. UgtP is required for β-lactam resistance in methicillin-resistant S. aureus (MRSA), and the inhibitors restored sensitivity to oxacillin in a highly resistant S. aureus strain. As no other compounds were pursued as possible LTA glycolipid assembly inhibitors, this work demonstrates the extraordinary efficiency of screens that exploit synthetic lethality to discover compounds that target specified pathways. The general approach should be applicable not only to other bacteria but also to eukaryotic cells.

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The Novel Membrane-Associated Auxiliary Factors AuxA and AuxB Modulate β-lactam Resistance in MRSA by stabilizing Lipoteichoic Acids

Cited by 27 publications

References 39 publications

SmdA is a novel cell morphology determinant in Staphylococcus aureus

SmdA is a novel cell morphology determinant in Staphylococcus aureus

Controlling biofilm and virulence properties of Gram-positive bacteria by targeting wall teichoic acid and lipoteichoic acid

Rapid Inhibitor Discovery by Exploiting Synthetic Lethality

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