1996 Fiscal Year Final Research Report Summary
Study on Expression of the pseudogene for Glucocerebrosidase
| Project/Area Number |
08044289
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| Research Category |
Grant-in-Aid for international Scientific Research
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| Allocation Type | Single-year Grants |
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| Section | Joint Research |
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| Research Institution | Tsu City College |
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Principal Investigator |
IMAI Katsuyuki Tsu City College, Department of Science for Living, Professor, 生活科学科, 教授 (60035425)
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| Co-Investigator(Kenkyū-buntansha) |
GINNS Edward National Institute of Health, Clinical Ne, Acting Chi
NAKAMURA Masahiko Osaka University, Institute for Protein Research, タンパク質研究所, 助手 (20172439)
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| Project Period (FY) |
1996
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| Keywords | Glucocerebrosidase / Pseudogene / Gaucher / Lysosome |
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| Research Abstract |
Glucocerebrosidase (GC) is a lysosomal enzyme. Genetic defects in the enzyme cause Gaucher' disease. Though several mutations in GC have been detected in patients' cells, a variety of clinical phenotypes have not causally been related to genotypes so far found. In collaborative studies on GC gene expression in a myeloid cell line, we found that a pseudogene is expressed in the cells and the transcript produced in vitro a 30 kDa protein. The present project aimed to get an insight into a physiological meaning of the pseudogene.
The pseudogene was found to be expressed ubiquitously not only in cell lines, but also in such normal cells as human placenta and bone marrow. PCR method used for the detection gave us only qualitative results. Quantitative analysis in various patients cells are needed. Both the normal and pseudotype genes obtained have longer non-coding sequences at 3' region. In the light of a recent finding that 3'non-coding region may have a regulatory function, we searched for the presence of this sequence by PCR and found the same sequence in some cells. Study on difference in efficiency of translation between GC genes with and without this longer sequence is under progress.
Targetting mechanism of GC to lysosomes is still not known. GC has been reported to become membrane bound after exit from endoplasmic reticulum. We examined binding to membrane prepararions of GC products made from plasmid constructs devoid of specific regions to identify a structure required for the specific binding. Only translation product from whole sequence was capable of the specific binding, suggesting a quaternary structure is necessary for the binding. Identification and cloning of a binding protein by use of yeast two hybrid system is under progress in collaboration with Dr.Ginns in NIH.
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