1887

Abstract

Phasins play an important role in the formation of poly(3-hydroxybutyrate) [poly(3HB)] granules and affect their size. Recently, three homologues of the phasin protein PhaP1 were identified in strain H16. The functions of PhaP2, PhaP3 and PhaP4 were examined by analysis of H16 deletion strains (Δ, Δ, Δ, Δ, Δ, Δ and Δ). When cells were grown under conditions permissive for poly(3HB) accumulation, the wild-type strain and all single-phasin negative mutants (Δ, Δ and Δ), with the exception of Δ, showed similar growth and poly(3HB) accumulation behaviour, and also the size and number of the granules were identical. The single Δ mutant and the Δ, Δ and Δ mutants showed an almost identical growth behaviour; however, they accumulated poly(3HB) at a significantly lower level than wild-type and the single Δ, Δ or Δ mutants. Gel-mobility-shift assays and DNaseI footprinting experiments demonstrated the capability of the transcriptional repressor PhaR to bind to a DNA region +36 to +46 bp downstream of the start codon. The protected sequence exhibited high similarity to the binding sites of PhaR upstream of , which were identified recently. In contrast, PhaR did not bind to the upstream or intergenic regions of and , thus indicating that the expression of these two phasins is regulated in a different way. Our current model for the regulation of phasins in strain H16 was extended and confirmed.

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2005-03-01
2019-08-21
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Appearance of confluently grown colonies of the various mutants in comparison to the wild-type, and gel-mobility-shift assays of PhaR binding to DNA fragments comprising up- and downstream regions of , and [PDF file](2108 KB)

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