PMC

Isolation of gain-of-susceptibility mutants of A. thaliana Col-0 by using the TEV-bar selectable virus. (A) Identification of mutants A130 and A158 as survivors after GA application. The photographs were taken 13 days after GA application. (B) Capsid protein (CP) immunoblot analysis of purified TEV capsid protein (lane 1), inflorescence tissue from a mock-inoculated C24 plant (lane 2), or inflorescence tissue from C24, Col-3, or GA survivors A130, A133, and A138 inoculated with TEV-bar (lanes 3–7). (C) β-Glucuronidase (GUS) activity assays of inflorescence tissue from M3 progeny of GA survivors (A130, A133, A138, A154, and A158) or restrictive (Col-3) and susceptible (C24) control plants inoculated with TEV-GUS. GUS assays were done at 15 days p.i. or 16 days p.i. for A154 and A158.

Results of the IPM-EDTA synergy test for carbapenemase-producing A. baumannii clinical isolates. (A) Imipenem disk (10 μg) alone. (B) Imipenem disk supplemented with 10 μl of EDTA at 0.5 M. A significant increase of the inhibition zone in the presence of EDTA was considered positive. From left to right, the panels contain an NDM-1-producing A. baumannii strain (isolate SLO), an NDM-2-producing A. baumannii strain (isolate ML), an IMP-4-producing A. baumannii strain (isolate IMP), a VIM-4-producing Acinetobacter genomospecies 16 strain (isolate A154), an OXA-23-producing A. baumannii strain (isolate Acb2), and an OXA-58-producing A. baumannii (isolate MAY).

(A) Ribbon representation of EGFP highlighting β-strands in green, helices in red and loops in pale green. Inset, Stick representation of the central helix forming residues (main chain only) in the 3₁₀ conformation (dark red) and in the α conformation (yellow). Residue Leu60 (orange) is involved in hydrogen bonding in the 3₁₀ portion of the helix (grey dashed lines) and the α portion of the helix (black dashed lines). (B) B-Factor putty representation of EGFP with chromophore shown as sticks. The B-factor color scale is shown to the right. The regions with high B-factors (shown as bulges) are: K3 to G10 (N-terminus); G115 to D116; K131 to G134; A154 to I161; P187 to P196; K209 to E213; G228 to L231 (C-terminus).

Modeling of the FabG(Ts) mutant proteins. (A) Tetrameric structure of FabG (29) with the monomer chains shown in different colors. The residues involved in the A154T, E233K, and S224F mutations are shown as balls and sticks. In each case the structures are shown in pairs (B plus C, D plus E, and F plus G) with the wild-type FabG structure to the left and the modeled mutant structure to the right. (B and C) Consequences of the E233K mutation. The salt bridge between E233 and H236′, indicated by a dashed red line in panel B, is missing in panel C due to the E233K mutation. (D) View of interactions around A154 at the other dimer interface of the protein. (E) Modeled replacement of A154 with Thr. The α5 helix was shortened due to the formation of a hydrogen bond (dashed green line) between Y151 and T154, thereby increasing the distance between α5 and α5′. (F and G) S224F mutation. Panel F is a view of the dimer interface interactions around Ser 224. Panel G suggests that replacement of Ser 224 with Phe could destabilize the stacking between Phe 221 and Phe 221′ due to loss of hydrogen bonding by the hydroxyl groups of Ser 224 and Ser 224′. This mutation could also disrupt hydrogen bonding between Ser 224 and Glu 226.