The role of the sulfur globule proteins of Allochromatium vinosum: mutagenesis of the sulfur globule protein genes and expression studies by real-time RT-PCR.
During oxidation of reduced sulfur compounds, the purple sulfur bacterium Allochromatium vinosum stores sulfur in the periplasm in the form of intracellular sulfur globules. The sulfur in the globules is enclosed by a protein envelope that consists of the homologous 10.5-kDa proteins SgpA and SgpB and the smaller 8.5-kDa SgpC. Reporter gene fusions of sgpA and alkaline phosphatase showed the constitutive expression of sgpA in A. vinosum and yielded additional evidence for the periplasmic localization of the sulfur globules. Expression analysis of the wild-type sgp genes by quantitative RT-PCR using the LightCycler system showed the constitutive expression of all three sgp genes. The expression of sgpB and sgpC is significantly enhanced under photolithotrophic conditions. Interestingly, sgpB is expressed ten times less than sgpA and sgpC implying that SgpA and SgpC are the "main proteins" of the sulfur globule envelope. Mutants with inactivated sgpA or sgpB did not show any differences in comparison with the wild-type, i.e., the encoded proteins can replace each other, whereas inactivation of sgpC leads to the formation of considerably smaller sulfur globules. This indicates a role of SgpC for globule expansion. A sgpBC double mutant was unable to grow on sulfide and could not form sulfur globules, showing that the protein envelope is indispensible for the formation and deposition of intracellular sulfur.
Prange
Alexander
A
Institut für Mikrobiologie & Biotechnologie, Rheinische Friedrich-Wilhelms-Universität Bonn, Meckenheimer Allee 168, 53115 Bonn, Germany.
Engelhardt
Harald
H
Trüper
Hans G
HG
Dahl
Christiane
C
D000469:Alkaline Phosphatase / Q000235:genetics / Q000378:metabolism; D019677:Artificial Gene Fusion; D001426:Bacterial Proteins / Q000235:genetics* / Q000502:physiology*; D002841:Chromatiaceae / Q000235:genetics* / Q000502:physiology* / Q000648:ultrastructure; D017353:Gene Deletion; D020869:Gene Expression Profiling; D017930:Genes, Reporter; D008970:Molecular Weight; D009154:Mutation; D033903:Periplasmic Proteins / Q000235:genetics / Q000502:physiology; D012329:RNA, Bacterial / Q000032:analysis / Q000302:isolation & purification; D012333:RNA, Messenger / Q000032:analysis / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013455:Sulfur / Q000378:metabolism*; D014158:Transcription, Genetic
D001426:Bacterial Proteins; D033903:Periplasmic Proteins; D012329:RNA, Bacterial; D012333:RNA, Messenger; C112535:SgpA protein, bacterial; C112537:SgpB protein, bacterial; C112538:SgpC protein, bacterial; D013455:Sulfur; D000469:Alkaline Phosphatase