Nucleoid-associated PhaF phasin drives intracellular location and segregation of polyhydroxyalkanoate granules in Pseudomonas putida KT2442

The PhaF is a nucleoid-associated like protein of Pseudomonas putida KT2442 involved in the polyhydroxyalkanoate (PHA) metabolism. Its primary structure shows two modular domains; the N-terminal PHA granule-binding domain (phasin domain) and the C-terminal half containing AAKP-like tandem repeats ch...

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Veröffentlicht in:Molecular microbiology 2011, Vol.79 (2), p.402-418
Hauptverfasser: Galán, B, Dinjaski, N, Maestro, B, de Eugenio, L.I, Escapa, I.F, Sanz, J.M, García, J.L, Prieto, M.A
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container_end_page 418
container_issue 2
container_start_page 402
container_title Molecular microbiology
container_volume 79
creator Galán, B
Dinjaski, N
Maestro, B
de Eugenio, L.I
Escapa, I.F
Sanz, J.M
García, J.L
Prieto, M.A
description The PhaF is a nucleoid-associated like protein of Pseudomonas putida KT2442 involved in the polyhydroxyalkanoate (PHA) metabolism. Its primary structure shows two modular domains; the N-terminal PHA granule-binding domain (phasin domain) and the C-terminal half containing AAKP-like tandem repeats characteristic of the histone H1 family. Although the PhaF binding to PHA granules and its role as transcriptional regulator have been previously demonstrated, the cell physiology meaning of these properties remains unknown. This work demonstrates that PhaF plays a crucial role in granule localization within the cell. TEM and flow cytometry studies of cells producing granules at early growth stage demonstrated that PhaF directs the PHA granules to the centre of the cells, forming a characteristic needle array. Our studies demonstrated the existence of two markedly different cell populations in the strain lacking PhaF protein, i.e. cells with and without PHA. Complementation studies definitively demonstrated a key role of PhaF in granule segregation during the cell division ensuring the equal distribution of granules between daughter cells. In vitro studies showed that PhaF binds DNA through its C-terminal domain in a non-specific manner. All these findings suggested a main role of PhaF in PHA apparatus through interactions with the segregating chromosome.
doi_str_mv 10.1111/j.1365-2958.2010.07450.x
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Its primary structure shows two modular domains; the N-terminal PHA granule-binding domain (phasin domain) and the C-terminal half containing AAKP-like tandem repeats characteristic of the histone H1 family. Although the PhaF binding to PHA granules and its role as transcriptional regulator have been previously demonstrated, the cell physiology meaning of these properties remains unknown. This work demonstrates that PhaF plays a crucial role in granule localization within the cell. TEM and flow cytometry studies of cells producing granules at early growth stage demonstrated that PhaF directs the PHA granules to the centre of the cells, forming a characteristic needle array. Our studies demonstrated the existence of two markedly different cell populations in the strain lacking PhaF protein, i.e. cells with and without PHA. 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Complementation studies definitively demonstrated a key role of PhaF in granule segregation during the cell division ensuring the equal distribution of granules between daughter cells. In vitro studies showed that PhaF binds DNA through its C-terminal domain in a non-specific manner. All these findings suggested a main role of PhaF in PHA apparatus through interactions with the segregating chromosome.</abstract><cop>Oxford, UK</cop><pub>Blackwell Publishing Ltd</pub><pmid>21219460</pmid><doi>10.1111/j.1365-2958.2010.07450.x</doi><tpages>17</tpages><oa>free_for_read</oa></addata></record>
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source Wiley-Blackwell Journals; Wiley-Blackwell Open Access Backfiles; MEDLINE; EZB Electronic Journals Library
subjects Bacteria
Bacterial Proteins - genetics
Bacterial Proteins - metabolism
Bacteriology
Binding sites
Biological and medical sciences
Carrier Proteins - genetics
Carrier Proteins - metabolism
Cells
Chromosomes
Cytoplasmic Granules - metabolism
Flow Cytometry
Fundamental and applied biological sciences. Psychology
Gene Deletion
Genetic Complementation Test
Metabolism
Microbiology
Microscopy, Electron, Transmission
Miscellaneous
Molecular biology
Polyhydroxyalkanoates - metabolism
Proteins
Pseudomonas putida
Pseudomonas putida - genetics
Pseudomonas putida - metabolism
title Nucleoid-associated PhaF phasin drives intracellular location and segregation of polyhydroxyalkanoate granules in Pseudomonas putida KT2442
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