Marques Lucas M, Rezende Izadora S, Barbosa Maysa S, Guimarães Ana M S, Martins Hellen B, Campos Guilherme B, do Nascimento Naíla C, Dos Santos Andrea P, Amorim Aline T, Santos Verena M, Farias Sávio T, Barrence Fernanda  C, de Souza Lauro M, Buzinhani Melissa, Arana-Chavez Victor E, Zenteno Maria E, Amarante-Mendes Gustavo P, Messick Joanne B, Timenetsky Jorge
Department of Microbiology, Institute of Biomedical Science, University of São Paulo, São Paulo, Brazil.
Multidisciplinary Institute of Health, Universidade Federal da Bahia, Vitória da Conquista, Brazil.
PLoS One. 2016 Sep 7;11(9):e0161926. doi: 10.1371/journal.pone.0161926. eCollection 2016.
Whole genome sequencing and analyses of Ureaplasma diversum ATCC 49782 was undertaken as a step towards understanding U. diversum biology and pathogenicity. The complete genome showed 973,501 bp in a single circular chromosome, with 28.2% of G+C content. A total of 782 coding DNA sequences (CDSs), and 6 rRNA and 32 tRNA genes were predicted and annotated. The metabolic pathways are identical to other human ureaplasmas, including the production of ATP via hydrolysis of the urea. Genes related to pathogenicity, such as urease, phospholipase, hemolysin, and a Mycoplasma Ig binding protein (MIB)-Mycoplasma Ig protease (MIP) system were identified. More interestingly, a large number of genes (n = 40) encoding surface molecules were annotated in the genome (lipoproteins, multiple-banded antigen like protein, membrane nuclease lipoprotein and variable surface antigens lipoprotein). In addition, a gene encoding glycosyltransferase was also found. This enzyme has been associated with the production of capsule in mycoplasmas and ureaplasma. We then sought to detect the presence of a capsule in this organism. A polysaccharide capsule from 11 to 17 nm of U. diversum was observed trough electron microscopy and using specific dyes. This structure contained arabinose, xylose, mannose, galactose and glucose. In order to understand the inflammatory response against these surface molecules, we evaluated the response of murine macrophages J774 against viable and non-viable U. diversum. As with viable bacteria, non-viable bacteria were capable of promoting a significant inflammatory response by activation of Toll like receptor 2 (TLR2), indicating that surface molecules are important for the activation of inflammatory response. Furthermore, a cascade of genes related to the inflammasome pathway of macrophages was also up-regulated during infection with viable organisms when compared to non-infected cells. In conclusion, U. diversum has a typical ureaplasma genome and metabolism, and its surface molecules, including the identified capsular material, represent major components of the organism immunopathogenesis.
对解脲脲原体ATCC 49782进行全基因组测序和分析,是迈向了解解脲脲原体生物学特性和致病性的重要一步。完整基因组显示,其单一环状染色体大小为973,501 bp,G+C含量为28.2%。共预测并注释了782个编码DNA序列(CDS)、6个rRNA基因和32个tRNA基因。其代谢途径与其他人类脲原体相同,包括通过尿素水解产生ATP。鉴定出了与致病性相关的基因,如脲酶、磷脂酶、溶血素以及支原体Ig结合蛋白(MIB)-支原体Ig蛋白酶(MIP)系统。更有趣的是,基因组中注释了大量编码表面分子的基因(n = 40)(脂蛋白、多带抗原样蛋白、膜核酸酶脂蛋白和可变表面抗原脂蛋白)。此外,还发现了一个编码糖基转移酶的基因。该酶与支原体和脲原体中荚膜的产生有关。随后,我们试图检测该生物体中是否存在荚膜。通过电子显微镜和使用特定染料,观察到了解脲脲原体11至17 nm的多糖荚膜。该结构含有阿拉伯糖、木糖、甘露糖、半乳糖和葡萄糖。为了了解针对这些表面分子的炎症反应,我们评估了小鼠巨噬细胞J774对活的和非活的解脲脲原体的反应。与活细菌一样,非活细菌能够通过激活Toll样受体2(TLR2)引发显著的炎症反应,表明表面分子对炎症反应的激活很重要。此外,与未感染细胞相比,在活生物体感染期间,与巨噬细胞炎性小体途径相关的一系列基因也会上调。总之,解脲脲原体具有典型的脲原体基因组和代谢,其表面分子,包括已鉴定的荚膜物质,是该生物体免疫发病机制的主要组成部分。