Phosphoethanolamine modification of lipid A in colistin-resistant variants of Acinetobacter baumannii mediated by the pmrAB two-component regulatory system

Colistin resistance is rare in Acinetobacter baumannii, and little is known about its mechanism. We investigated the role of PmrCAB in this trait, using (i) resistant and susceptible clinical strains, (ii) laboratory-selected mutants of the type strain ATCC 19606 and of the clinical isolate ABRIM, and (iii) a susceptible/resistant pair of isogenic clinical isolates, Ab15/133 and Ab15/132, isolated from the same patient. pmrAB sequences in all the colistin-susceptible isolates were identical to reference sequences, whereas resistant clinical isolates harbored one or two amino acid replacements variously located in PmrB. Single substitutions in PmrB were also found in resistant mutants of strains ATCC 19606 and ABRIM and in the resistant clinical isolate Ab15/132. No mutations in PmrA or PmrC were found. Reverse transcriptase (RT)-PCR identified increased expression of pmrA (4- to 13-fold), pmrB (2- to 7-fold), and pmrC (1- to 3-fold) in resistant versus susceptible organisms. Matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry showed the addition of phosphoethanolamine to the hepta-acylated form of lipid A in the resistant variants and in strain ATCC 19606 grown under low-Mg induction conditions. pmrB gene knockout mutants of the colistin-resistant ATCC 19606 derivative showed >100-fold increased susceptibility to colistin and 5-fold decreased expression of pmrC; they also lacked the addition of phosphoethanolamine to lipid A. We conclude that the development of a moderate level of colistin resistance in A. baumannii requires distinct genetic events, including (i) at least one point mutation in pmrB, (ii) upregulation of pmrAB, and (iii) expression of pmrC, which lead to addition of phosphoethanolamine to lipid A. Copyright © 2011, American Society for Microbiology. All Rights Reserved.

Transit timing variations in WASP-10b induced by stellar activity

The hot-JupiterWASP-10bwas reported by Maciejewski et al. to showtransit timing variations (TTVs) with an amplitude of ~3.5 min. These authors proposed that the observed TTVs were caused by a 0.1M_Jup perturbing companion with an orbital period of ~5.23 d, and hence, close to the outer 5:3 mean-motion resonance with WASP-10b. To test this scenario, we present eight new transit light curves of WASP-10b obtained with the Faulkes Telescope North and the Liverpool Telescope. The new light curves, together with 22 previously published ones, were modelled with a Markov Chain Monte Carlo transit fitting code. Transit depth differences reported forWASP-10b are thought to be due to starspot-induced brightness modulation of the host star. Assuming the star is brighter at the activity minimum, we favour a small planetary radius. We find Rp = 1.039^+0.043 _-0.049R_Jup in agreement with Johnson et al. and Maciejewski et al. Recent studies find no evidence for a significant eccentricity in this system. We present consistent system parameters for a circular orbit and refine the orbital ephemeris ofWASP-10b. Our homogeneously derived transit times do not support the previous claimed TTV signal, which was strongly dependent on two previously published transits that have been incorrectly normalized. Nevertheless, a linear ephemeris is not a statistically good fit to the transit times of WASP-10b. We show that the observed transit time variations are due to spot occultation features or systematics. We discuss and exemplify the effects of occultation spot features in the measured transit times and show that despite spot occultation during egress and ingress being difficult to distinguish in the transit light curves, they have a significant effect in the measured transit times. We conclude that if we account for spot features, the transit times of WASP-10b are consistent with a linear ephemeris with the exception of one transit (epoch 143) which is a partial transit. Therefore, there is currently no evidence for the existence of a companion to WASP-10b. Our results support the lack of TTVs of hot-Jupiters reported for the Kepler sample.