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논문 기본 정보
- 자료유형
- 학술저널
- 저자정보
- 발행연도
- 2003.1
- 수록면
- 393 - 402 (10page)
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We report here the isolation, characterization on
genomic structure and expression of the D. melanogaster
homolog of human parkin. The 2,122 bp
parkin gene sequence contains six exons that
form a 1,449 bp transcript encoding a protein of
482 amino acids. 151 bp of 5' and 112 bp of 3'
untranslated regions were identified by a combination
of 5'-RACE/primer extension and 3'-RACE,
respectively. The 5' UTR contains three transcription
initiation sites. Neither a classical TATA nor
a CAAT box was found in the putative promoter
sequence. However, binding sites for AhR-Arnt,
AP4, NF1 and GATA transcription factors were
identified. Transient transfection analysis of the 5'
UTR confirmed its promoter activity in HEK 293
cells and SH-SY5Y neuronal cells using a dual luciferase
reporting system. The amino acid sequence
of D. melanogaster Parkin exhibits 42%,
43% and 43% identity to that of human, mouse
and rat, respectively, representing a 54 kDa protein
band via western blot analysis. It shows a
high degree of conservation in the Ubiquitin-like
domain at the N-terminus (34%), the In-Between
RING finger domains (IBR, 65-69%), and the RING
finger domains at the C-terminus (56-57%). The
expression pattern of D. melanogaster parkin
varies during the developmental stages, with the
highest expression in the adult stage as measured
by competitive RT-PCR. From immunostainings of
the embryo, D. melanogaster parkin was expressed
slightly higher in the central nervous system
(brain and nerve cord) during the late embryonic
stage.
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