Publikationsserver der RWTH Aachen University
Subzelluläre Lokalisation des Y-box-Proteins-1 (YB-1) in Mesangialzellen und Charakterisierung der dafür verantwortlichen Proteindomänen
Abstract
dc:descriptionAs a DNA- and RNA binding protein the cold-shock protein YB-1 serves multiple functions in the nucleus and cytoplasm, e.g. regulating transcription, RNA-processing, -translation and –storage. In the present study we examined the expression and localisation pattern of YB-1 in various models of glomerular diseases. Furthermore we determined protein regions involved in the regulation of YB-1 localisation. In glomeruli of healthy rat kidneys YB-1 is detected exclusively in the nucleus by means of immunohistochemistry with two different antibodies against YB-1. In the course of the Ant-Thy 1.1- nephritis YB-1 changes its localisation and expression pattern. Within one day YB-1 localizes to the perinuclear region of cells within all glomeruli . In the course of the disease the strongest cytoplasmic YB-1 signal coincided with the peak mesangial cell activation/proliferation between days 4 and 6 and was detected in a mesangial distribution pattern. With resolution of the disease at day 31 with complete morphological restitution was associated with YB-1 relocalisation to the nucleus. In the course of different other models of rat glomerular disease YB-1 remained nuclear. In the second part of the study we examined YB-1 localisation in the in vitro model of proliferating rat mesangial cells, which are activated in cell culture conditions. In these cells endogenous YB-1 as well as transfected GFP-YB-1-proteins was detected in the cytoplasm. Using cell cycle inhibitors we tried to arrest the rat mesangial cells. Only after treatment with roscovitine, a specific CDK- inhibitor, YB-1 translocated into the nucleus. Cloning various deletion construct of YB-1 with a fluorescent tag we identified three nuclear localisation signals (NLS: AS 185-194, AS 149-156, AS 276-292) and two cytoplasmic localisation signals or nuclear export signals ( ZLS: AS 1-57, AS 52-146). Experiments using leptomycin B as an inhibitor of the CRM1 dependent nuclear export could reveal a CRM-1 independent export of YB-1. In summary these results document an increased expression and translocation of YB-1 in mesangial cells in the course of glomerulonephritis, therefore YB-1 has pivotal role coordinating inflammatory processes in glomerular disease.
Degree
thesis:*- Grantor dc:publisher
- Publikationsserver der RWTH Aachen University
- Year dc:date
- 2008
Author and committee
dc:creator, dc:contributor.*- Author dc:creator
-
- Martinkus, Sandra
- Contributors dc:contributor
-
- Mertens, Peter René
Subjects
dc:subject × 13Rights
dc:rights- Statement dc:rights
-
- info:eu-repo/semantics/openAccess
- Language dc:language
- ger