| Project/Area Number |
16390315
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (B)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
Dermatology
|
|---|
| Research Institution | Nagoya University |
|---|
Principal Investigator |
TOMITA Yasushi Graduate School of Medicine, Professor, 大学院医学系研究科, 教授 (70108512)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
SUZUKI Tamio Yamagata University School of Medicine, Professor, 医学部, 教授 (30206502)
YASUE Takashi University Hospital, Research Associate, 医学部附属病院, 助手 (40335039)
|
|---|
| Project Period (FY) |
2004 – 2006
|
| Project Status |
Completed (Fiscal Year 2006)
|
| Budget Amount *help |
¥14,500,000 (Direct Cost: ¥14,500,000)
Fiscal Year 2006: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 2005: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 2004: ¥11,900,000 (Direct Cost: ¥11,900,000)
|
|---|
| Keywords | adenosine deaminase / Dyschromatosis / ADAR1 / DSRAD / RNA editing |
|---|
| Research Abstract |
Dyschromatosis symmmetrica hereditaria (DSH) is a pigmentary disorder with an autosomal dominant inheritance. In 2003, we for the first time reported that the double-stranded RNA-specific adenosine deaminase (DSRAD/ADAR1) is the responsible gene for DSH. ADAR1 protein catalyzes the deamination of adenosine to inosine in double-stranded RNA substrates, which results in the creation of alternative splicing sites or alternations of the codon and thus leads to functional changes in the protein. Purpose of this study was to clarify the pathomechanism how DSH developed in the patient with the gene mutation of ADAR1.
(1) Human ADAR1 protein has been reported to include two major forms, one is the interferon-inducible full-length 150-kDa protein (p150) and the other is the constitutively expressed N-terminally truncated 110-kDa protein (p110). The production of the two forms is due to the presence of different promoters. In this study, we have found two mutations causing frameshift changes in t
… More
he synthesis of p150, but an effect on the synthesis of p110 protein. Thus we can suggest that the full-length p150 must be involved with DSH.
(2) In order to identify the RNA substrate for p150ADAR1, we tried to screen out several candidates as the substrate(s) using two strategies as follows: (a) Oligonucleotide array analyses of interferon-inducible p150ADAR1 genes in cultured melanocytes. As the p150ADAR1 is induced by interferon, and transfection with siADAR1 was reported to dramatically inhibite the expression of ADAR1, we compared the total cellular RNA expression in normal human melanocytes cultured with interferon and that with the short inhibitory mRNA (siRNAs) of p150ADAR1 by analyses using the human gene arrays. (b) Computational identification of RNAs with A-to-I editing sites. In principle, editing can be detected using the large-scale database of expressed sequence tags (ESTs) and RNAs. Editing sites show up when a sequence is aligned with the genome: while the DNA reads A, sequencing identifies the inosine in the edited site as guanosine.
As the results of two experiments as mentioned above, we listed many candidate RNAs, but have identified no specific RNA substrate(s) among them. Less
|
