Differences in mitochondrial genome organization of Cryptococcus neoformans strains.

The organization of the mitochondrial genomes in two strains belonging in different varieties of Cryptococcus neoformans was analysed. Physical maps of the mtDNA of the IFM5844 (var. neoformans) and IFO410 (var. grubii) strains were constructed by using EcoRI and EcoRV restriction enzymes; functional maps were constructed by hybridization, cloning and sequencing. Most of the genes important in the mitochondrial function (ND1, ND2, ND3, ND4, ND4L, ND5, ND6, ATP6, ATP9, COX1, COX2 and COB) and protein synthesis (SsrRNA and LsrRNA) were localized. We did not find any differences between the strains in the order of these genes. However, they differed significantly in the sizes of the mtDNAs: 32.6 kb for IFM5844, and 24.1 kb for IFO410. This can be attributed to two large regions of the mtDNA. In these regions, differences were found in the numbers of introns in COX1 (no intron in var. grubii, 5 introns in var. neoformans), COB (1 intron in var. grubii, 2 introns in var. neoformans), LsrRNA (no intron in var. grubii, 2 introns in var. neoformans), and ND5 (no intron in var. grubii, 1 intron in var. neoformans) genes. In several introns of the COB and COX1 genes LAGLIDADG motifs were found. Differences were also observed in the nucleotide sequences of some genes and in the sizes and sequences of intergenic regions. The nucleotide sequences of the genes of the IFM and IFO strains were compared with those of the H-99 and JEC 21 strains from the database. Surprisingly high similarities were found between the strains belonging in var. grubii (IFO 410 and H-99) and var. neoformans (IFM 5844 and JEC 21).

Organization of mitochondrial DNA in the basidiomycetous Dioszegia hungarica (Cryptococcus hungaricus) species.

The organization of mitochondrial DNA was investigated in the six collection strains of the basidiomycetous yeast Dioszegia hungarica (Cryptococcus hungaricus) isolated so far. Physical and partial functional maps were constructed. Two strains (CBS 6324 and 6576) were identical while three others (CBS 4214, 5124, 6953) differed not only in the distribution of restriction sites but in gene order as well. Results confirm the hypothesis that these five strains are representatives of different closely related species. The sixth strain CBS 6569 revealed a unique mitochondrial genome organization. Its mtDNA separated into eight bands on agarose gel without enzymatic digestion. These molecules carried mitochondrial genes, and RFLP analysis of the four largest molecules using frequently-cutting restriction enzymes (KpnI and SmaI) showed them to have strongly homologous sequences. This unique mtDNA organization was also observed in a strain of Cystofilobasidium capitatum, providing evidence that CBS 6569 belongs to the Cystofilobasidium clade.