A prophage-encoded actin-like protein required for efficient viral DNA replication in bacteria.
In host cells, viral replication is localized at specific subcellular sites. Viruses that infect eukaryotic and prokaryotic cells often use host-derived cytoskeletal structures, such as the actin skeleton, for intracellular positioning. Here, we describe that a prophage, CGP3, integrated into the genome of Corynebacterium glutamicum encodes an actin-like protein, AlpC. Biochemical characterization confirms that AlpC is a bona fide actin-like protein and cell biological analysis shows that AlpC forms filamentous structures upon prophage induction. The co-transcribed adaptor protein, AlpA, binds to a consensus sequence in the upstream promoter region of the alpAC operon and also interacts with AlpC, thus connecting circular phage DNA to the actin-like filaments. Transcriptome analysis revealed that alpA and alpC are among the early induced genes upon excision of the CGP3 prophage. Furthermore, qPCR analysis of mutant strains revealed that both AlpA and AlpC are required for efficient phage replication. Altogether, these data emphasize that AlpAC are crucial for the spatio-temporal organization of efficient viral replication. This is remarkably similar to actin-assisted membrane localization of eukaryotic viruses that use the actin cytoskeleton to concentrate virus particles at the egress sites and provides a link of evolutionary conserved interactions between intracellular virus transport and actin.
Department of Biology I, Ludwig-Maximilians-University Munich, Großhaderner Str. 2-4, 82152 Planegg-Martinsried, Germany Institute for Biochemistry, University of Cologne, Zülpicherstr. 47, 50674 Cologne, Germany.
Catriona
C
Donovan
Institut für Bio- und Geowissenschaften, IBG-1: Biotechnologie, Forschungszentrum Jülich, D-52425 Jülich, Germany.
Antonia
A
Heyer
Institut für Bio- und Geowissenschaften, IBG-1: Biotechnologie, Forschungszentrum Jülich, D-52425 Jülich, Germany.
Eugen
E
Pfeifer
Institut für Bio- und Geowissenschaften, IBG-1: Biotechnologie, Forschungszentrum Jülich, D-52425 Jülich, Germany.
Tino
T
Polen
Institute for Biochemistry, University of Cologne, Zülpicherstr. 47, 50674 Cologne, Germany.
Anja
A
Wittmann
Institute for Biochemistry, University of Cologne, Zülpicherstr. 47, 50674 Cologne, Germany.
Reinhard
R
Krämer
Institut für Bio- und Geowissenschaften, IBG-1: Biotechnologie, Forschungszentrum Jülich, D-52425 Jülich, Germany j.frunzke@fz-juelich.de.
Julia
J
Frunzke
Department of Biology I, Ludwig-Maximilians-University Munich, Großhaderner Str. 2-4, 82152 Planegg-Martinsried, Germany Institute for Biochemistry, University of Cologne, Zülpicherstr. 47, 50674 Cologne, Germany marc.bramkamp@lmu.de.
Marc
M
Bramkamp
D000199:Actins / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure; D000255:Adenosine Triphosphate / Q000378:metabolism; D048230:Corynebacterium glutamicum / Q000235:genetics / Q000821:virology*; D004261:DNA Replication; D004279:DNA, Viral / Q000032:analysis / Q000096:biosynthesis*; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D006160:Guanosine Triphosphate / Q000378:metabolism; D039002:Prophages / Q000235:genetics* / Q000502:physiology; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure; D014779:Virus Replication
D000199:Actins; D004279:DNA, Viral; D004268:DNA-Binding Proteins; D014764:Viral Proteins; D006160:Guanosine Triphosphate; D000255:Adenosine Triphosphate