Cloning of a centromere binding factor 3d (CBF3D) gene fromCandida glabrata

Stoyan, Tanja; Eck, Raimund; Lechner, Johannes; Hemmerich, Peter; Künkel, W.; Diekmann, Stephan

doi:10.1002/(sici)1097-0061(19990630)15:9<793::aid-yea415>3.0.co;2-g

Cited by 3 publications

(4 citation statements)

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“…Thus, it was of interest to determine whether the inner kinetochore protein homologs are species specific. Based on the ability of an Scskp1 deletion strain containing CgSKP1 on a LEU2-bearing plasmid to grow on 5-FOA (which means that the ScSKP1 gene on a URA3-bearing plasmid can be lost), CgSkp1p was previously shown to complement an Scskp1⌬ mutant (60). To determine whether the other C. glabrata inner kinetochore proteins could complement deletions of the corresponding genes in S. cerevisiae, we inserted the C. glabrata genes under the control of their own promoters into plasmids that contained either an ScLEU2 or an ScHIS3 marker and tested them in the corresponding S. cerevisiae deletion strains (plasmids pRSCg1, pTS32, pTS39, pTS37, pRSCg3D, and pTS34) ( Table 1; see also Table S1 in the supplemental material for plasmid constructs).…”

Section: Resultsmentioning

confidence: 99%

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Inner Kinetochore of the Pathogenic YeastCandida glabrata

Stoyan

Carbon

2004

Eukaryot Cell

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The human pathogenic yeast Candida glabrata is the second most common Candida pathogen after Candida albicans, causing both bloodstream and mucosal infections. The centromere (CEN) DNA of C. glabrata (CgCEN), although structurally very similar to that of Saccharomyces cerevisiae, is not functional in S. cerevisiae. To further examine the structure of the C. glabrata inner kinetochore, we isolated several C. glabrata homologs of S. cerevisiae inner kinetochore protein genes, namely, genes for components of the CBF3 complex (Ndc10p, Cep3p, and Ctf13p) and genes for the proteins Mif2p and Cse4p. The amino acid sequence identities of these proteins were 32 to 49% relative to S. cerevisiae. CgNDC10, CgCEP3, and CgCTF13 are required for growth in C. glabrata and are specifically found at CgCEN, as demonstrated by chromatin immunoprecipitation experiments. Cross-complementation experiments revealed that the isolated genes, with the exception of CgCSE4, are species specific and cannot functionally substitute for the corresponding genes in S. cerevisiae deletion strains. Likewise, the S. cerevisiae CBF3 genes NDC10, CEP3, and CTF13 cannot functionally replace their homologs in C. glabrata CBF3 deletion strains. Two-hybrid analysis revealed several interactions between these proteins, all of which were previously reported for the inner kinetochore proteins of S. cerevisiae. Our findings indicate that although many of the inner kinetochore components have evolved considerably between the two closely related species, the organization of the C. glabrata inner kinetochore is similar to that in S. cerevisiae.

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Section: Resultsmentioning

confidence: 99%

“…In previous work, two of the inner kinetochore proteins of C. glabrata, Skp1p and Cbf1p (CgSkp1p and CgCbf1p, respectively), were cloned and characterized (60,61). CgSkp1p is 76% identical to and functionally exchangeable with the S. cerevisiae homolog.…”

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Inner Kinetochore of the Pathogenic YeastCandida glabrata

Stoyan

Carbon

2004

Eukaryot Cell

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“…The multiprotein complex Cbf3 binds to CDEIII and consists of the proteins Cbf3a, Cbf3b, Cbf3c and Cbf3d (Stemmann and Lechner, 1996). The only centromeric protein that was cloned from human pathogenic yeasts is Cbf3d from C. glabrata (Stoyan et al, 1999).…”

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confidence: 99%

The centromere‐binding factor Cbf1p from Candida albicans complements the methionine auxotrophic phenotype of Saccharomyces cerevisiae

Eck

Stoyan

Künkel

2001

Yeast

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A gene encoding the centromere binding factor 1 (Cbf1p) of the human pathogenic yeast Candida albicans was cloned and characterized. An open reading-frame was detected which encoded a 223 amino acid protein with a calculated molecular weight of 25.8 kDa and a relative isoelectric point of 5.55. It shares 39% overall amino acid sequence identity with Saccharomyces cerevisiae Cbf1p. We localized the CaCBF1 gene on chromosome 4. Southern analysis indicated that CaCBF1 is probably present as a single copy gene per haploid genome. The CaCBF1 gene under the control of its own promoter was able to complement the methionine auxotrophic growth, the increased mitotic instability of CEN plasmids and the slow growth of a Saccharomyces cerevisiae cbf1D mutant strain. The

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“…Recently, we have cloned a CBF3d homolog from C. glabrata and showed that it can functionally substitute for its S. cerevisiae counterpart (57). In this study we present the cloning and functional characterization of the CBF1 gene of C. glabrata (CgCBF1).…”

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