Segmental conservation of sapA sequences in type B Campylobacter fetus cells

Abstract

Campylobacter fetus cells may exist as either of two defined serogroups (type A or B) based on their lipopolysaccharide (LPS) composition. Wild-type strains contain surface array proteins (S-layer proteins) that have partial antigenic cross-reactivity but bind exclusively to LPS from homologous (type A or B) cells. Type A cells possess 8 homologs of sapA, which encodes a 97-kDa S-layer protein; the gene products of these homologs have a conserved N terminus of 184 amino acids. To further explore the structural relationships between the C. fetus S-layer proteins and their encoding genes, we sought to clone and express an S-layer protein from type B strain 84-91. The cloned type B gene (sapB) was similar in structure to the previously cloned type A gene (sapA) and encoded a full-length 936-amino acid (97-kDa) S-layer protein. Sequence analysis of sapB indicated that the conserved N-terminal encoding region in sapA was absent but that the remainder of the ORF (encoding 751 amino acids) was identical to that of sapA in spite of the nonconserved nature of this region among sapA homologs. Noncoding sequences both 300 base pairs 5' and 1000 base pairs 3' to the sapB and sapA ORFs, including the sapA promoter and transcriptional terminator sequences, were essentially identical. Southern analyses revealed that the sapB N-terminal encoding region was conserved in multiple copies in type B strains but was absent in type A strains. Recombinant sapA and sapB products bound to a substantially greater degree to cells of the homologous LPS type compared with the heterologous LPS type, indicating that the conserved sapA- and sapB-encoded N termini are critical for LPS binding specificity. The parallel genetic organization and identity at the nucleotide level in both coding and noncoding regions for sap homologs in types A and B cells indicates the necessity of both homolog conservation and high fidelity DNA replication in the biology of sap diversity.