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Volume 152, Issue 5

Nisin induction without nisin secretion Free

Abstract

Nisin Z, a post-translationally modified antimicrobial peptide of , is positively autoregulated by extracellular nisin via the two-component regulatory proteins NisRK. A mutation in the nisin NisT transporter rendered incapable of nisin secretion, and nisin accumulated inside the cells. Normally nisin is activated after secretion by the serine protease NisP in the cell wall. This study showed that when secretion of nisin was blocked, intracellular proteolytic activity could cleave the N-terminal leader peptide of nisin precursor, resulting in active nisin. The isolated cytoplasm of a non-nisin producer could also cleave the leader from the nisin precursor, showing that the cytoplasm of cells does contain proteolytic activity capable of cleaving the leader from fully modified nisin precursor. Nisin could not be detected in the growth supernatant of the NisT mutant strain with a nisin-sensing strain (sensitivity 10 pg ml), which has a green fluorescent protein gene connected to the nisin-inducible promoter and a functional nisin signal transduction circuit. Northern analysis of the NisT mutant cells revealed that even though the cells could not secrete nisin, the nisin-inducible promoter P was active. In a or background, where nisin could not be fully modified due to the mutations in the nisin modification machinery, the unmodified or partly modified nisin precursor accumulated in the cytoplasm. This immature nisin could not induce the P promoter. The results suggest that when active nisin is accumulated in the cytoplasm, it can insert into the membrane and from there extrude parts of the molecule into the pseudoperiplasmic space to interact with the signal-recognition domain of the histidine kinase NisK. Potentially, signal presentation via the membrane represents a general pathway for amphiphilic signals to interact with their sensors for signal transduction.

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Keyword(s): GFP, green fluorescent protein , HPK, histidine protein kinase , RFU, relative fluorescence unit , RR, response regulator and TCST, two-component signal transduction
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2006-05-01
2024-04-23
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Nisin induction without nisin secretion
Microbiology 152, 1489 (2006); https://doi.org/10.1099/mic.0.28544-0
/content/journal/micro/10.1099/mic.0.28544-0
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