UMR INRA 1014 Sécurité des aliments et Microbiologie, Ecole Nationale Vétérinaire, 44307 Nantes, France.
Appl Environ Microbiol. 2009 Dec;75(23):7310-8. doi: 10.1128/AEM.01366-09. Epub 2009 Oct 2.
Campylobacter jejuni is a leading cause of bacterial gastroenteritis worldwide. Infection generally occurs after ingestion of contaminated poultry products, usually conserved at low temperatures. The mechanisms promoting survival of C. jejuni in the cold remain poorly understood despite several investigations. The present study provides insight into the survival mechanism by establishing the involvement of polynucleotide phosphorylase (PNPase), a 3'-5' exoribonuclease with multiple biological functions in cold survival. The role of PNPase was demonstrated genetically using strains with altered pnp genes (which encode PNPase) created in C. jejuni F38011 and C. jejuni 81-76 backgrounds. Survival assays carried out at low temperatures (4 and 10 degrees C) revealed a difference of 3 log CFU/ml between the wild-type and the pnp deletion (Deltapnp) strains. This did not result from a general requirement for PNPase because survival rates of the strains were similar at higher growth temperatures (37 or 42 degrees C). trans-Complementation with plasmid pNH04 carrying the pnp gene under the control of its natural promoter restored the cold survival phenotype to the pnp deletion strains (at 4 and 10 degrees C) but not to the same level as the wild type. In this study we demonstrate the role of PNPase in low-temperature survival of C. jejuni and therefore attribute a novel biological function to PNPase directly related to human health.
空肠弯曲菌是全球细菌性胃肠炎的主要病因。感染通常发生在摄入受污染的家禽产品后,这些产品通常在低温下保存。尽管已经进行了多次调查,但促进空肠弯曲菌在低温下存活的机制仍知之甚少。本研究通过建立多核苷酸磷酸化酶(PNPase)在冷适应中发挥作用的机制,为冷适应中的存活机制提供了新的认识。多核苷酸磷酸化酶(PNPase)是一种具有多种生物学功能的 3'-5'外切核糖核酸酶,在 C. jejuni F38011 和 C. jejuni 81-76 背景下用改变了的 pnp 基因(编码 PNPase)的菌株进行遗传研究,从而证明了 PNPase 的作用。在低温(4 和 10°C)下进行的存活实验表明,野生型和 pnp 缺失(Deltapnp)菌株之间相差 3 个对数 CFU/ml。这不是由于 PNPase 的一般要求造成的,因为在较高的生长温度(37 或 42°C)下,菌株的存活率相似。在其天然启动子控制下携带 pnp 基因的质粒 pNH04 的反式互补恢复了 pnp 缺失菌株(在 4 和 10°C)的低温存活表型,但没有恢复到野生型的水平。在这项研究中,我们证明了 PNPase 在空肠弯曲菌低温存活中的作用,因此将 PNPase 的一个新的生物学功能直接归因于与人类健康相关的功能。