RESUMEN
By transformation of a Saccharomyces cerevisiae mutant strain conditionally expressing the ABF1-gene, a Kluyveromyces marxianus DNA fragment carrying the gene encoding the ABF1-homologue of this yeast strain (KmABF1) was selected. Comparison of the sequence of the KmABF1 gene with that encoding Saccharomyces cerevisiae ABF1 and the previously isolated ABF1-gene from Kluyveromyces lactis (KlABF1) revealed distinct regions displaying considerable homology and therefore most likely representing sequences encoding essential domains. In addition to the domains putatively involved in DNA binding of the protein factor, two short conserved amino acid sequence elements at the C-termini of the homologous proteins were identified, which are proposed to play a part in their trans-acting functions. This is the first report on the structure of a regulatory protein factor from K. marxianus.
Asunto(s)
Proteínas de Unión al ADN/química , Proteínas Fúngicas/química , Kluyveromyces/química , Proteínas de Saccharomyces cerevisiae , Factores de Transcripción/química , Secuencia de Aminoácidos , Proteínas de Unión al ADN/genética , Proteínas Fúngicas/genética , Prueba de Complementación Genética , Kluyveromyces/genética , Datos de Secuencia Molecular , Saccharomyces cerevisiae/genética , Homología de Secuencia de Aminoácido , Factores de Transcripción/genéticaRESUMEN
Recently, a mutant fibronectin gene was identified in skin fibroblasts obtained from sclerotic lesions of 7 Japanese patients with systemic sclerosis (SSc). Two point mutations were found adjacent to the cell-attachment tetrapeptide DNA sequence in exon 7 of the fibronectin gene. In the present study, we investigated whether these point mutations are present in the fibronectin gene of Dutch patients with SSc. We were unable to demonstrate the point mutations in the Dutch SSc patients studied.
Asunto(s)
Fibronectinas/genética , Genes , Mutación , Esclerodermia Sistémica/genética , Secuencia de Bases , Exones , Humanos , Japón , Datos de Secuencia Molecular , Países Bajos , Reacción en Cadena de la PolimerasaRESUMEN
An enzyme-linked immunosorbent assay (ELISA) has been developed for the detection of anticentromere autoantibodies in sera of patients with suspected or manifest rheumatic diseases. The antigen source used in this assay consists of the recombinant protein of glutathione S-transferase (GST) fused to the last 60 C-terminal amino acid residues of the major centromere protein CENP-B. Although this CENP-B segment is only a small part of the complete polypeptide, we show that it constitutes an important autoimmune antigenic domain which is recognized by all patient sera in which ACA can be detected using the immunoblotting technique with a HeLa S3 nuclear protein extract as antigen source.