含組氨酸的磷酸轉移蛋白B (HptB, PA3345)是在Pseudomonas aeruginosa PAO1中，參與將磷酸基從多重感應激酶轉移到反應調控子PA3346的其中之一個蛋白。本論文研究目標是探討HptB-PA3346交互作用和其調控機制的生物意義。剔除hptB基因突變株作轉錄分析(transcription-profiling analysis)後，發現一系列與移動性相關的基因改變，與突變株群泳(swarming)性狀的觀察情形一致。蛋白質結構分析發現，PA3346的C端區域(PA3346C)與枯草桿菌 anti-sigma因子SpoIIAB顯示出大約百分之三十的相似度。利用體外磷酸化分析得知，Ser/Thr蛋白激酶活化anti-sigma拮抗劑PA3347胺基酸Ser-56，此現象也在PA3346C中被證實。此外，PA3346C和anti-σ28因子FlgM發現不管在體內還是體外實驗，都與PA3347有直接作用。在反應時，FlgM取代PA3346C與PA3347結合，之後σ28完全取代掉PA3347-FlgM複合體中的 PA3347，形成磷酸根依賴性夥伴交換系統。藉由分析剔除PA3346基因突變株和其互補株，本研究建立PA3347磷酸化在連結夥伴交換系統和群泳移動力中，所扮演的重要性。在hptB基因突變株將磷酸酯解酶過度表現，則會發現群泳性狀缺陷的情況會回復，顯示出htpB蛋白調節細胞內訊號分子環狀雙鳥嘌呤單磷酸(cyclic-di-GMP)訊息傳遞路徑進而調控群泳性狀。

The Histidine-containing phosphotransfer protein-B (HptB, PA3345) is an intermediate protein involved in transferring a phosphoryl group from multiple sensor kinases to the response regulator PA3346 in Pseudomonas aeruginosa PAO1. The objective of this study is to elucidate the biological significance of the HptB-PA3346 interaction and the regulatory mechanisms thereafter. The transcription-profiling analysis of an hptB knockout mutant showed that the expression of a number of motility-related genes was altered, consistent with the non-swarming phenotype observed for the mutant. Domain analysis indicates that the PA3346 C-terminal region (PA3346C) exhibits approximately 30% identity with the anti-sigma factor SpoIIAB of Bacillus subtilis. The presence of Ser/Thr protein kinase activity-targeting an anti-sigma antagonist PA3347 at Ser-56 was confirmed in PA3346C using an in vitro phosphorelay assay. Furthermore, PA3346C and the anti-σ28 factor FlgM were found to interact with PA3347 individually both in vivo and in vitro. FlgM displaced PA3346C in binding of PA3347 and was then competitively displaced by σ28 from the PA3347-FlgM complex, forming a phosphorylation-dependent partner-switching system. The significance of PA3347 phosphorylation in linking the partner-switching system and swarming motility was established by analyzing the swarming phenotype of the PA3347 knockout mutant and its complement strain. The over expression of phosphodiesterase in hptB mutant restore its swarming motility defect showing the role of intracellular signaling molecule cyclic-di-GMP in regulating the motility behavior of HptB mediated signaling pathway.

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