Utility of in vivo transcription profiling for identifying Pseudomonas aeruginosa genes needed for gastrointestinal colonization and dissemination.
Utility of in vivo transcription profiling for identifying Pseudomonas aeruginosa genes needed for gastrointestinal colonization and dissemination.
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DOI：
10.1371/journal.pone.0015131
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发表时间：
2010-12-10
期刊：
影响因子：
3.7
通讯作者：
Pier GB
中科院分区：
文献类型：
--
作者：
Koh AY;Mikkelsen PJ;Smith RS;Coggshall KT;Kamei A;Givskov M;Lory S;Pier GB
以前，对动物感染期间在体内表达的铜绿假单胞菌mRNA转录本的微阵列分析尚未被用于调查这一环境所需的潜在毒力因子。我们比较了从铜绿假单胞菌定植的小鼠的胃肠道(GI)中回收的细菌细胞中的mRNA表达，以及用于定植小鼠的饮用水中的铜绿假单胞菌。与生物膜形成和III型分泌相关的基因(T3SS)在胃肠道中的表达显著增加。铜绿假单胞菌PA14菌株中的非冗余转座子文库被用来测试在体内定植过程中被鉴定为转录增加的基因的突变体。所有生物膜相关基因的Tn文库突变体在体外形成生物膜的能力都有所减弱，但这些突变体在胃肠道的定植和扩散方面与WT铜绿假单胞菌PA14没有显著差异。为了评估T3SS因子，我们测试了在铜绿假单胞菌T3SS、ExOS或ExoU、ExoT和POPB的四个基因中，GI定植和中性粒细胞减少导致的缺失(PAO1和PAK)和插入(PA14)突变的传播。这些突变株与其对应的WT株之间在胃肠道定植方面没有显著差异，而感染T3SS突变株的小鼠传播后存活率显著下降。然而，亲本突变体和T3SS突变体之间的总体存活率存在可变的、依赖于菌株的影响。因此，体内小鼠胃肠道定植期间基因转录的增加并不能预测基因产物在中性粒细胞减少症诱导后的定植或总体存活中的重要作用。
Microarray analysis of Pseudomonas aeruginosa mRNA transcripts expressed in vivo during animal infection has not been previously used to investigate potential virulence factors needed in this setting. We compared mRNA expression in bacterial cells recovered from the gastrointestinal (GI) tracts of P. aeruginosa-colonized mice to that of P. aeruginosa in the drinking water used to colonize the mice. Genes associated with biofilm formation and type III secretion (T3SS) had markedly increased expression in the GI tract. A non-redundant transposon library in P. aeruginosa strain PA14 was used to test mutants in genes identified as having increased transcription during in vivo colonization. All of the Tn-library mutants in biofilm-associated genes had an attenuated ability to form biofilms in vitro, but there were no significant differences in GI colonization and dissemination between these mutants and WT P. aeruginosa PA14. To evaluate T3SS factors, we tested GI colonization and neutropenia-induced dissemination of both deletional (PAO1 and PAK) and insertional (PA14) mutants in four genes in the P. aeruginosa T3SS, exoS or exoU, exoT, and popB. There were no significant differences in GI colonization among these mutant strains and their WT counterparts, whereas rates of survival following dissemination were significantly decreased in mice infected by the T3SS mutant strains. However, there was a variable, strain-dependent effect on overall survival between parental and T3SS mutants. Thus, increased transcription of genes during in vivo murine GI colonization is not predictive of an essential role for the gene product in either colonization or overall survival following induction of neutropenia.
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影响因子：
3.1
作者：
Koh, AY;Priebe, GP;Pier, GB
通讯作者：
Pier, GB
DOI：
10.1152/ajpgi.2001.280.5.g922
发表时间：
2001-05-01
影响因子：
4.5
作者：
Atuma, C;Strugala, V;Holm, L
通讯作者：
Holm, L
DOI：
10.1038/nrmicro2199
发表时间：
2009-09
期刊：
Nature reviews. Microbiology
影响因子：
--
作者：
通讯作者：
--
影响因子：
3.1
作者：
Allewelt, M;Coleman, FT;Pier, GB
通讯作者：
Pier, GB
影响因子：
3.1
作者：
Alexander, RJ;Raicht, RF
通讯作者：
Raicht, RF