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Biology subjects

Curtis, E. R.

Publications and source records attributed to Curtis, E. R..

2 recordsLinked to original sources

Profiling Myxococcus xanthus swarming phenotypes through mutation and environmental variation

Myxococcus xanthus is a bacterium that lives on surfaces as a predatory biofilm called a swarm.As a growing swarm feeds on prey and expands, it displays dynamic multicellular patterns such as traveling waves called ripples and branching protrusions called flares. The rate at which a swarm expands across a surface, and the emergence of the coexisting patterns, are all controlled through coordinated cell movement. M. xanthus cells move using two motility systems known as Adventurous (A) and Social (S). Both are involved in swarm expansion and pattern formation. In this study, we describe a set of M. xanthus swarming genotype-to-phenotype associations that include both genetic and environmental perturbations. We identified new features of the swarming phenotype; recorded and measured swarm expansion using time-lapse microscopy; and compared the impact of mutation on different surfaces. These observations and analyses have increased our ability to discriminate between swarming phenotypes and provided context that allowed us to identify some phenotypes as improbable outliers within the M. xanthus swarming phenome. ImportanceMyxococcus xanthus grows on surfaces as a predatory biofilm called a swarm. A feeding swarm expands by moving over and consuming prey bacteria. In the laboratory, a swarm is created by spotting cell suspension onto nutrient agar in lieu of prey. The cells quickly settle on the surface and the new swarm then expands radially. An assay that measures the expansion rate of a swarm of mutant cells is the first, and sometimes only, measurement used to decide whether a particular mutation impacts swarm motility. We have broadened the scope of this assay by increasing the accuracy of measurements and reintroducing prey, resulting in new identifiable and quantifiable features that can be used to improve genotype-to-phenotype associations.

microbiology↗

Ghrelin O-acyltransferase interacts with extracellular peptides and exhibits unexpected cellular localization for a secretory pathway enzyme

Ghrelin O-acyltransferase (GOAT) plays a central role in the maturation and activation of the peptide hormone ghrelin, which performs a wide range of endocrinological signaling roles. Using a tight-binding fluorescent ghrelin-derived peptide designed for high selectivity for GOAT over the ghrelin receptor GHS-R1a, we demonstrate that GOAT interacts with extracellular ghrelin and facilitates ligand cell internalization in both transfected cells and prostate cancer cells endogenously expressing GOAT. Coupled with enzyme mutagenesis, ligand uptake studies provide the first direct evidence supporting interaction of the putative histidine general base within GOAT with the ghrelin peptide acylation site. Our work provides a new understanding of GOATs catalytic mechanism, establishes a key step required for autocrine/paracrine ghrelin signaling involving local reacylation by GOAT, and raises the possibility that other peptide hormones may exhibit similar complexity in their intercellular and organismal-level signaling pathways.

biochemistry↗