Janisch Niklas, Levendosky Keith, Budell William C, Quadri Luis E N
Department of Biology, Brooklyn College, City University of New York, 2900 Bedford Avenue, Brooklyn, NY 11210, USA.
Biology Program, Graduate Center, City University of New York, 365 Fifth Avenue, New York, NY 10016, USA.
Pathogens. 2023 Jan 5;12(1):86. doi: 10.3390/pathogens12010086.
() causes opportunistic pulmonary infections with tuberculosis-like features. The bacterium is well known for its photochromogenicity, i.e., the production of carotenoid pigments in response to light. The genetics defining the photochromogenic phenotype of has not been investigated and defined pigmentation mutants to facilitate studies on the role of carotenes in the bacterium's biology are not available thus far. In this study, we set out to identify genetic determinants involved in photochromogenicity. We screened a library of ~150,000 transposon mutants for colonies with pigmentation abnormalities. The screen rendered a collection of ~200 mutants. Each of these mutants could be assigned to one of four distinct phenotypic groups. The insertion sites in the mutant collection clustered in three chromosomal regions. A combination of phenotypic analysis, sequence bioinformatics, and gene expression studies linked these regions to carotene biosynthesis, carotene degradation, and monounsaturated fatty acid biosynthesis. Furthermore, introduction of the identified carotenoid biosynthetic gene cluster into non-pigmented endowed the bacterium with photochromogenicity. The studies also led to identification of MarR-type and TetR/AcrR-type regulators controlling photochromogenicity and carotenoid breakdown, respectively. Lastly, the work presented also provides a first insight into the transcriptome changes in response to light.
()会引发具有结核样特征的机会性肺部感染。这种细菌以其光致变色性而闻名,即响应光照产生类胡萝卜素色素。定义该菌光致变色表型的遗传学尚未得到研究,目前也没有用于促进研究类胡萝卜素在该菌生物学中作用的色素沉着突变体。在本研究中,我们着手鉴定参与该菌光致变色性的遗传决定因素。我们筛选了一个约150,000个转座子突变体文库,以寻找色素沉着异常的菌落。筛选得到了约200个突变体。这些突变体中的每一个都可被归为四个不同表型组之一。突变体文库中的插入位点聚集在三个染色体区域。通过表型分析、序列生物信息学和基因表达研究相结合,将这些区域与类胡萝卜素生物合成、类胡萝卜素降解和单不饱和脂肪酸生物合成联系起来。此外,将鉴定出的类胡萝卜素生物合成基因簇导入无色素的该菌中,使其具有光致变色性。这些研究还分别鉴定出了控制光致变色性和类胡萝卜素分解的MarR型和TetR/AcrR型调节因子。最后,所展示的工作还首次深入了解了该菌响应光照时的转录组变化。