1999 Fiscal Year Final Research Report Summary
Functional analysis of left-right determining (inv) gene
| Project/Area Number |
10680689
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Developmental biology
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| Research Institution | Tokyo Women's Medical University |
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Principal Investigator |
YOKOYAMA Takahiko Tokyo Women's Medical University, Department of Anatomy and Developmental Biology Joshu, 医学部, 助手 (70191525)
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| Project Period (FY) |
1998 – 1999
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| Keywords | left / right asymmetry / inv / situs inversus / ankyrin / intracellular localization |
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| Research Abstract |
Almost all internal organs of vertebrate body have distinctive left/right asymmetry such as heart and spleen on the left. The inv mutant shows a constant reversal of such left/right polarity. We have identified the gene that is responsible for the inv mutation. The gene encodes 1062 amino acids and 15 repeats of ankyrin motif and two bipartite nuclear localization signals. The gene has not previously reported and its cellular function is unknown. To study the intracellular function of the inv gene, we determined the localization of the inv gene product. GFP (green fluorescent protein) fusion constructs linked to the full length inv gene are generated and are transfected in cos 7 cells. Curiously, signals are observed in the cytoplasm when GFP is connected in front of the inv gene whereas in nucleus when GFP is connectd behind the inv gene. When flag tag or myc tag is connected in front of or behind the inv gene, signals are observed in the cytoplasm. Therefore we concluded that the inv protein is normally localized in the cytoplasm. We next deleted the inv gene and connected behind GFP gene. These constructs are also transfected in cos 7 cells and their localization was examined. When GFP constructs have bipartite nuclear localization signal without ankyrin repeats, signal are observed in the nucleus. Therefore, nuclear localization signals in the inv gene are functional. If the constructs retained ankyrin repeats, signals were observed in the cytoplasm but not in the nucleus. Although it is still unclear where the inv gene product works, these results suggested that the inv protein may be function by transporting between the nucleus and the cytoplasm.
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Research Products
(4 results)
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[Publications] T Mochizuki, Y Saijoh, K Tsuchiya, Y Shirayoshi, S Takai, C Taya, H Yonekawa, K Yamada, H Nihei, N Nakatsuji, PA Overbeek, H Hamada, T Yokoyama: "Cloning of inv, a gene that controls left/right asymmetry and kidney development."Nature. 395. 177-181 (1998)
Description
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