Localization pattern of conjugation machinery in a Gram-positive bacterium.
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Localization pattern of conjugation machinery in a Gram-positive bacterium. / Bauer, Theresa; Rösch, Thomas; Itaya, Mitsuhiro; Graumann, Peter L.
In: J BACTERIOL, Vol. 193, No. 22, 22, 2011, p. 6244-6256.Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review
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TY - JOUR
T1 - Localization pattern of conjugation machinery in a Gram-positive bacterium.
AU - Bauer, Theresa
AU - Rösch, Thomas
AU - Itaya, Mitsuhiro
AU - Graumann, Peter L
PY - 2011
Y1 - 2011
N2 - Conjugation is an efficient way for transfer of genetic information between bacteria, even between highly diverged species, and a major cause for the spreading of resistance genes. We have investigated the subcellular localization of several conserved conjugation proteins carried on plasmid pLS20 found in Bacillus subtilis. We show that VirB1, VirB4, VirB11, VirD2, and VirD4 homologs assemble at a single cell pole, but also at other sites along the cell membrane, in cells during the lag phase of growth. Bimolecular fluorescence complementation analyses showed that VirB4 and VirD4 interact at the cell pole and, less frequently, at other sites along the membrane. VirB1 and VirB11 also colocalized at the cell pole. Total internal reflection fluorescence microscopy showed that pLS20 is largely membrane associated and is frequently found at the cell pole, indicating that transfer takes place at the pole, which is a preferred site for the assembly of the active conjugation apparatus, but not the sole site. VirD2, VirB4, and VirD4 started to localize to the pole or the membrane in stationary-phase cells, and VirB1 and VirB11 were observed as foci in cells resuspended in fresh medium but no longer in cells that had entered exponential growth, although at least VirB4 was still expressed. These data reveal an unusual assembly/disassembly timing for the pLS20 conjugation machinery and suggest that specific localization of conjugation proteins in lag-phase cells and delocalization during growth are the reasons why pLS20 conjugation occurs only during early exponential phase.
AB - Conjugation is an efficient way for transfer of genetic information between bacteria, even between highly diverged species, and a major cause for the spreading of resistance genes. We have investigated the subcellular localization of several conserved conjugation proteins carried on plasmid pLS20 found in Bacillus subtilis. We show that VirB1, VirB4, VirB11, VirD2, and VirD4 homologs assemble at a single cell pole, but also at other sites along the cell membrane, in cells during the lag phase of growth. Bimolecular fluorescence complementation analyses showed that VirB4 and VirD4 interact at the cell pole and, less frequently, at other sites along the membrane. VirB1 and VirB11 also colocalized at the cell pole. Total internal reflection fluorescence microscopy showed that pLS20 is largely membrane associated and is frequently found at the cell pole, indicating that transfer takes place at the pole, which is a preferred site for the assembly of the active conjugation apparatus, but not the sole site. VirD2, VirB4, and VirD4 started to localize to the pole or the membrane in stationary-phase cells, and VirB1 and VirB11 were observed as foci in cells resuspended in fresh medium but no longer in cells that had entered exponential growth, although at least VirB4 was still expressed. These data reveal an unusual assembly/disassembly timing for the pLS20 conjugation machinery and suggest that specific localization of conjugation proteins in lag-phase cells and delocalization during growth are the reasons why pLS20 conjugation occurs only during early exponential phase.
KW - Protein Transport
KW - Bacillus subtilis/genetics/growth & development/metabolism
KW - Bacterial Proteins/genetics/metabolism
KW - Conjugation, Genetic
KW - Plasmids/genetics/metabolism
KW - Protein Transport
KW - Bacillus subtilis/genetics/growth & development/metabolism
KW - Bacterial Proteins/genetics/metabolism
KW - Conjugation, Genetic
KW - Plasmids/genetics/metabolism
M3 - SCORING: Journal article
VL - 193
SP - 6244
EP - 6256
JO - J BACTERIOL
JF - J BACTERIOL
SN - 0021-9193
IS - 22
M1 - 22
ER -