Comparative secretome analysis suggests low plant cell wall degrading capacity in Frankia symbionts.
BACKGROUND
Frankia sp. strains, the nitrogen-fixing facultative endosymbionts of actinorhizal plants, have long been proposed to secrete hydrolytic enzymes such as cellulases, pectinases, and proteases that may contribute to plant root penetration and formation of symbiotic root nodules. These or other secreted proteins might logically be involved in the as yet unknown molecular interactions between Frankia and their host plants. We compared the genome-based secretomes of three Frankia strains representing diverse host specificities. Signal peptide detection algorithms were used to predict the individual secretomes of each strain, and the set of secreted proteins shared among the strains, termed the core Frankia secretome. Proteins in the core secretome may be involved in the actinorhizal symbiosis.


RESULTS
The Frankia genomes have conserved Sec (general secretory) and Tat (twin arginine translocase) secretion systems. The potential secretome of each Frankia strain comprised 4-5% of the total proteome, a lower percentage than that found in the genomes of other actinobacteria, legume endosymbionts, and plant pathogens. Hydrolytic enzymes made up only a small fraction of the total number of predicted secreted proteins in each strain. Surprisingly, polysaccharide-degrading enzymes were few in number, especially in strain CcI3, with more esterolytic, lipolytic and proteolytic enzymes having signal peptides. A total of 161 orthologous proteins belong to the core Frankia secretome. Of these, 52 also lack homologs in closely related actinobacteria, and are termed "Frankia-specific." The genes encoding these conserved secreted proteins are often clustered near secretion machinery genes.


CONCLUSIONS
The predicted secretomes of Frankia sp. are relatively small and include few hydrolases, which could reflect adaptation to a symbiotic lifestyle. There are no well-conserved secreted polysaccharide-degrading enzymes present in all three Frankia genomes, suggesting that plant cell wall polysaccharide degradation may not be crucial to root infection, or that this degradation varies among strains. We hypothesize that the relative lack of secreted polysaccharide-degrading enzymes in Frankia reflects a strategy used by these bacteria to avoid eliciting host defense responses. The esterases, lipases, and proteases found in the core Frankia secretome might facilitate hyphal penetration through the cell wall, release carbon sources, or modify chemical signals. The core secretome also includes extracellular solute-binding proteins and Frankia-specific hypothetical proteins that may enable the actinorhizal symbiosis.
Mastronunzio
Juliana E
JE
Department of Molecular and Cell Biology, U-3125, University of Connecticut, Storrs, CT, USA. juliana.mastronunzio@uconn.edu
Tisa
Louis S
LS
Normand
Philippe
P
Benson
David R
DR
D002473:Cell Wall / Q000382:microbiology; D002874:Chromosome Mapping; D040161:Frankia / Q000235:genetics* / Q000502:physiology*; D016680:Genome, Bacterial; D006867:Hydrolases / Q000235:genetics / Q000502:physiology; D010802:Phylogeny; D010944:Plants / Q000382:microbiology*; D020543:Proteome; D013045:Species Specificity; D013559:Symbiosis / Q000235:genetics / Q000502:physiology
D020543:Proteome; D006867:Hydrolases