Molecular characterization of nitrite reductase gene (aniA) and gene product in Neisseria meningitidis isolates: is aniA essential for meningococcal survival?
The present study evaluates sequence conservation in the gene coding for nitrite reductase (aniA) and AniA expression from a panel of Neisseria meningitidis isolates. Sequence analysis of the coding region in 19 disease-associated and 4 carrier strains notwithstanding a high degree of sequence similarity showed a number of nucleotide changes, some of which possibly resulted in premature translation termination or function loss. In particular, in one disease-associated strain a 9-residues insertion was found to be located close to the type I Cu-site and a catalytic histidine at position 280 was mutated into a leucine. In two strains from carriers, a sequence corresponding to a portion of a transposase gene within the aniA was also found. The AniA protein was always expressed, except for these two carriers strains and for other two strains in which the presence of the premature stop codons was recognized. The biochemical properties of the cloned soluble domain of the enzyme (sAniA) from N. meningitidis reference MC58 strain and from a clinical invasive isolate were studied. In particular, biochemical analysis of sAniA from MC58 demonstrated a clear dependence of its catalytic activity upon acidification, while the clinical isolate-derived sAniA was not functional. Thus, the results obtained suggest that the presence of a conserved and functional aniA gene is not essential for meningococcal survival.
Stefanelli
Paola
P
Deptartment of Infectious, Parasitic and Immune-mediated diseases, Istituto Superiore di Sanità, Rome, Italy.
Colotti
Gianni
G
Neri
Arianna
A
Salucci
Maria Luisa
ML
Miccoli
Roberto
R
Di Leandro
Luana
L
Ippoliti
Rodolfo
R
D000328:Adult; D000595:Amino Acid Sequence; D000942:Antigens, Bacterial / Q000235:genetics* / Q000378:metabolism; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics* / Q000378:metabolism; D002648:Child; D003001:Cloning, Molecular; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D009345:Neisseria meningitidis / Q000201:enzymology / Q000235:genetics; D009572:Nitrite Reductases / Q000235:genetics* / Q000378:metabolism; D009573:Nitrites / Q000378:metabolism; D016415:Sequence Alignment; D020539:Sequence Analysis, Protein
D000942:Antigens, Bacterial; D001425:Bacterial Outer Membrane Proteins; D009573:Nitrites; D009572:Nitrite Reductases