A combination of cell biological, molecular biological and biochemical analyses was used to study a novel member of ClipR family, CLIPR76. Sequence analysis and experimental evidence show that the CLIPR76 gene is alternatively spliced and produces multiple isoforms in different organisms. Western blot and real time PCR investigations show that the CLIPR76 gene is ubiquitously expressed with highest levels in testis and striated muscle. Some isoforms appear to be tissue specific. Immunofluorescence microscopy of the GFP tagged or untagged CLIPR76 proteins demonstrate that the different isoforms have different cellular localization and imply that they have different protein activities. Some CLIPR76 isoforms localize to the endoplasmic reticulum and their overexpression affect the morphology of ER and ERGIC membrane compartments, while others localize to microtubules or subsets of microtubules and cause alterations in microtubule organization when expressed at higher levels. These results imply a role for CLIPR76 proteins in membrane transport and ER microtubules interactions.Figure 3.2 Schematic of BP recombination reaction The BP reaction is catalyzed by BP clonase mix (Invitrogen). Diagram taken from Gateway technology manual (Invitrogen). CLIPR76 MT1 CLIPR76 MT3 (CLIPR76 C120a). 43 1 Schematic ofanbsp;...
|Title||:||Characterization of a Novel Microtubule Binding Protein CLIPR76|
|Author||:||Gergana T. Ugrinova|
|Publisher||:||ProQuest - 2007|