Morphological changes and proteome response of Corynebacterium glutamicum to a partial depletion of FtsI.
In Corynebacterium glutamicum, as in many Gram-positive bacteria, the cell division gene ftsI is located at the beginning of the dcw cluster, which comprises cell division- and cell wall-related genes. Transcriptional analysis of the cluster revealed that ftsI is transcribed as part of a polycistronic mRNA, which includes at least mraZ, mraW, ftsL, ftsI and murE, from a promoter that is located upstream of mraZ. ftsI appears also to be expressed from a minor promoter that is located in the intergenic ftsL-ftsI region. It is an essential gene in C. glutamicum, and a reduced expression of ftsI leads to the formation of larger and filamentous cells. A translational GFP-FtsI fusion protein was found to be functional and localized to the mid-cell of a growing bacterium, providing evidence of its role in cell division in C. glutamicum. This study involving proteomic analysis (using 2D SDS-PAGE) of a C. glutamicum strain that has partially depleted levels of FtsI reveals that at least 20 different proteins were overexpressed in the organism. Eight of these overexpressed proteins, which include DivIVA, were identified by MALDI-TOF. Overexpression of DivIVA was confirmed by Western blotting using anti-DivIVA antibodies, and also by fluorescence microscopy analysis of a C. glutamicum RESF1 strain expressing a chromosomal copy of a divIVA-gfp transcriptional fusion. Overexpression of DivIVA was not observed when FtsI was inhibited by cephalexin treatment or by partial depletion of FtsZ.
Valbuena
Noelia
N
Departamento de Ecología, Genética y Microbiología, Área de Microbiología, Facultad de Biología, Universidad de León, 24071 León, Spain.
Letek
Michal
M
Departamento de Ecología, Genética y Microbiología, Área de Microbiología, Facultad de Biología, Universidad de León, 24071 León, Spain.
Ramos
Angelina
A
Departamento de Ecología, Genética y Microbiología, Área de Microbiología, Facultad de Biología, Universidad de León, 24071 León, Spain.
Ayala
Juan
J
Centro Biología Molecular 'Severo Ochoa', Consejo Superior de Investigaciones Científicas, CSIC-UAM, Campus Universidad Autónoma, Cantoblanco, 28049 Madrid, Spain.
Nakunst
Diana
D
Institut fur Genomforschung, Universitat Bielefeld, Universitatsstrasse 25, D-33615 Bielefeld, Germany.
Kalinowski
Joern
J
Institut fur Genomforschung, Universitat Bielefeld, Universitatsstrasse 25, D-33615 Bielefeld, Germany.
Mateos
Luis M
LM
Departamento de Ecología, Genética y Microbiología, Área de Microbiología, Facultad de Biología, Universidad de León, 24071 León, Spain.
Gil
José A
JA
Departamento de Ecología, Genética y Microbiología, Área de Microbiología, Facultad de Biología, Universidad de León, 24071 León, Spain.
D001426:Bacterial Proteins / Q000032:analysis* / Q000235:genetics; D001483:Base Sequence; D018797:Cell Cycle Proteins / Q000032:analysis* / Q000235:genetics; D002506:Cephalexin / Q000494:pharmacology; D048230:Corynebacterium glutamicum / Q000737:chemistry / Q000166:cytology* / Q000235:genetics; D008969:Molecular Sequence Data; D046915:Penicillin-Binding Proteins / Q000502:physiology*; D020543:Proteome; D011993:Recombinant Fusion Proteins / Q000032:analysis
D001426:Bacterial Proteins; D018797:Cell Cycle Proteins; C104636:DivIVA protein, bacteria; D046915:Penicillin-Binding Proteins; D020543:Proteome; D011993:Recombinant Fusion Proteins; D002506:Cephalexin