Fig. 5: Inactivation of facZ impairs cell division and is rescued by deletion of gpsB. | Nature Microbiology

Fig. 5: Inactivation of facZ impairs cell division and is rescued by deletion of gpsB.

From: FacZ is a GpsB-interacting protein that prevents aberrant division-site placement in Staphylococcus aureus

Fig. 5

a, Transposon insertion profiles of the ezrA locus in strains aTB015 [WT] and aTB259 [∆facZ]. b, Spot titre of cultures of aTB003 [WT], aTB372 [∆facZ Ptet-facZ], aTP481 [∆ezrA Ptet-ezrA] and aTB378 [∆facZ ∆ezrA Ptet-facZ]. Cells were normalized to OD600 = 1.0, serially diluted and spotted on TSA agar with or without an aTC inducer (50 ng ml−1). c, Spot titre of aTB003 [WT], aTB372 [∆facZ Ptet-facZ] and aTP481 [∆ezrA Ptet-ezrA] as in b except that the plates contained PC190723 (100 ng ml−1). d, Top: Diagram showing the location of mutations in gpsB that suppress the PC190723 sensitivity of a ∆facZ mutant. Suppressor mutations are mapped onto a diagram of the two folded domains of GpsB (lines indicate position of mutations, with red lines indicating mutations generating premature stop codons). Bottom: Cultures of strains aTB003 [WT], aTB251 [∆facZ], aTB453 [∆facZ gpsB-s1], aTB476 [∆facZ gpsB-s2], aTB478 [∆facZ gpsB-s3] and aTB497 [∆facZ gpsB::Tn] were OD normalized and spotted on TSA supplemented with PC190723 (100 ng ml−1) as in b. e, Representative fluorescence images of aTB003 [WT], aTB372 [∆facZ Ptet-facZ], aTB525 [∆gpsB] and aTB540 [∆gpsB ∆facZ Ptet-facZ]. Strains were grown to mid-log phase without induction of facZ, and membranes were stained with Nile red (Extended Data Fig. 6). Yellow arrowheads highlight membrane defects. Scale bar = 2 µm. The scale bar applies to all images in e. f,g, Cultures of aTB521 [WT], aTB527 [∆facZ], aTB529 [∆gpsB] and aTB542 [∆facZ ∆gpsB] constitutively expressing cytoplasmic red fluorescent protein from pKK30 were labelled with TMA–DPH in mid-log phase and imaged on 2% agarose pads. f, Violin plots showing the cell area of indicated strains based on cytoplasmic fluorescence (WT, n = 1,760 cells; ∆facZ, n = 701 cells; ∆gpsB, n = 1,544 cells; ∆facZ ∆gpsB, n = 1,217 cells). Cell shape was quantified with MicrobeJ (Methods). g, The number of aberrant membrane foci per cell was quantified for display in pie charts (WT, n = 791 cells; ∆facZ, n = 400 cells; ∆gpsB, n = 998 cells; ∆facZ ∆gpsB, n = 721 cells).

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