Publikationsserver der RWTH Aachen University
Identifizierung und Untersuchung eines Magnesium-Transportproteins in der Spalthefe Schizosaccharomyces pombe
Abstract
dc:descriptionThe protein VMU1p (Vacuolar magnesium utilization) is a member of the MIT family (metal ion transporter) and has domains homologous to CorA-like Mg2+ transport proteins. This protein is encoded by Schizosaccharomyces pombe reading frame O13779. VMU1p contains the highly conserved motive GMN, which was found in all known magnesium transport proteins. These results support the idea that VMLU1p is a magnesium transporter.In order to identify the function of VMU1p, the vmu gen of the diploid S. pombe strain 130x131 was disrupted. The correct integration was controlled by PCR and Southern blotting. The tetrad analyses demonstrated a 2+:2- segregation of the his3 selection marker. This shows that VMU1p is not essential. Under normal grow conditions the disruptant and the wildtype strains showed similar growth rates. If the cells grew in medium without magnesium, the disrupted strains stopped growing earlier compared to the wildtype. The addition of magnesium restored growth. A magnesium specific staining showed that magnesium remained in the vacuoles in the disruptant strain but not in the wild type. These study suggests that magnesium transport into the cell takes place both in the disrupted and the wildtype strain, but magnesium mobilisation out of the vacuole within the cell is affected in the disruptant.Transformation of the disruptant with the VMU1 reading frame complemented the phenotype.Expression of a VMU1p-GFP fusion protein in S. pombe showed that VMU1p is localized in the vacuole membrane.
Degree
thesis:*- Grantor dc:publisher
- Publikationsserver der RWTH Aachen University
- Year dc:date
- 2006
Author and committee
dc:creator, dc:contributor.*- Author dc:creator
-
- Hammelmann, Silke
- Contributors dc:contributor
-
- Wolf, Klaus
Subjects
dc:subject × 5Rights
dc:rights- Statement dc:rights
-
- info:eu-repo/semantics/openAccess
- Language dc:language
- ger
Identifiers
dc:identifier.*- OAI identifier oai:identifier
- oai:publications.rwth-aachen.de:60969