The scc spirochetal coiled-coil protein forms helix-like filaments and binds to nucleic acids generating nucleoprotein structures.
The analysis of the genome of Leptospira spp., a group of bacteria of the phylum of spirochetes with several unique evolutionary and morphological features, has allowed the identification of a gene encoding a coiled-coil protein, called Scc, which is completely unrelated to any other eukaryotic or prokaryotic protein. Since coiled-coil proteins are often key elements of the cytoskeleton, we analyzed the protein Scc, which is a 24-kDa protein composed of a N-terminal coiled-coil domain, a proline-rich intermediate domain, and an acidic tail. The gene scc is located in an operon which also contains the genes encoding the initiation factor IF3 and the two ribosomal proteins L20 and L35. In this study, we showed that the presence of the coiled-coil domain was responsible for the polymerization of Scc in helix-like structures, in an ATP-independent manner, in both Escherichia coli living cells and in vitro. Analysis of the Scc polymers by electron microscopy showed filaments with a width of 6 to 10 nm, similar to that of eukaryotic intermediate filaments. Scc was also found to bind both RNA and double-stranded DNA without detectable sequence specificity. By electron microscopy, we showed that Scc polymer assembly was affected by the presence of nucleic acids, giving rise to rod-shaped structures with a width ranging from 45 to 155 nm. Finally, Leptospira biflexa cells depleted in Scc form small colonies, but the morphology of their helicoidal cell body was not affected. These results provide the first insight into a unique DNA binding filament-forming coiled-coil protein that could play an important role in the subcellular architecture of the spirochetal microorganism.
Mazouni
Khalil
K
Laboratoire des Spirochètes, Institut Pasteur, 28 rue du docteur Roux, 75724 Paris Cedex 15, France.
Pehau-Arnaudet
Gérard
G
England
Patrick
P
Bourhy
Pascale
P
Saint Girons
Isabelle
I
Picardeau
Mathieu
M
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D004269:DNA, Bacterial / Q000378:metabolism; D005798:Genes, Bacterial; D007382:Intermediate Filaments / Q000378:metabolism* / Q000648:ultrastructure; D007919:Leptospira / Q000737:chemistry* / Q000235:genetics / Q000648:ultrastructure; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009698:Nucleoproteins / Q000737:chemistry / Q000378:metabolism*; D011108:Polymers; D011392:Proline; D020836:Protein Structure, Quaternary; D017434:Protein Structure, Tertiary; D012329:RNA, Bacterial / Q000378:metabolism; D016415:Sequence Alignment
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D009698:Nucleoproteins; D011108:Polymers; D012329:RNA, Bacterial; D011392:Proline