Staphylococcus aureus ArlRS is a key two-component regulatory system necessary for adhesion, biofilm formation, and virulence. The response regulator ArlR consists of a C-terminal DNA-binding effector domain and an N-terminal receiver domain that is phosphorylated by ArlS, the cognate transmembrane sensor histidine kinase. We demonstrate that the receiver domain of ArlR adopts the canonical α5β5 response regulator assembly, which dimerizes upon activation, using beryllium trifluoride as an aspartate phosphorylation mimic. Activated ArlR recognizes a 20-bp imperfect inverted repeat sequence in the ica operon, which is involved in intercellular adhesion polysaccharide production. Crystal structures of the inactive and activated forms reveal that activation induces a significant conformational change in the β4-α4 and β5-α5-connecting loops, in which the α4 and α5 helices constitute the homodimerization interface. Crystal structures of the DNA-binding ArlR effector domain indicate that it is able to dimerize via a non-canonical β1–β2 hairpin domain swapping, raising the possibility of a new mechanism for signal transduction from the receiver domain to effector domain. Taken together, the current study provides structural insights into the activation of ArlR and its recognition, adding to the diversity of response regulation mechanisms that may inspire novel antimicrobial strategies specifically targeting Staphylococcus.
A soil urostylid ciliate Extraholosticha sylvatica, isolated from southern China, is studied using live observation and protargol impregnation. The main ontogenetic features of E. sylvatica are as follows: (a) in the proter, only the posterior part of the parental adoral zone of membranelles is renewed, while the anterior portion is retained; undulating membranes anlage undergo, like most hypotrichs, depolymerization of old undulating membranes and differentiation of new structures; (b) the oral primordium in the opisthe and the frontoventraltransverse cirral anlagen in both daughter cells are formed with contribution of the parental midventral cirri; (c) three frontal, one buccal, three to five frontoventral, one or two frontoterminal, two or three pretransverse ventral and six to 11 transverse cirri and 16–22 midventral pairs are generated from frontoventral-transverse cirral anlagen; (d) in both the left and the right marginal rows as well as the dorsal kineties, each two anlagen are formed intrakinetally; three to six caudal cirri are formed at the posterior end of the 5th dorsal kinety anlage; (e) the macronuclear nodules fuse into a single mass during the process.
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