9 research outputs found
Analysis of mDRA (Mouse Down-Regulated in Adenoma)-PP2A Bฮด interaction in the yeast two-hybrid system
/๋ฐ์ฌ[ํ๊ธ]
[์๋ฌธ]
The expressed recombinant cDNA of mouse Down-Regulated in Adenoma (mDRA) originally proposed as a candidate tumor suppressor mediates DIDS-sensitive, Na^+-independent, electroneutral Cl^-/HCOโ^- exchange (34). Several potential
regulatory sites of nuclear localization signals and numerous putative phosphorylation sites within the COOH-terminus of the deduced amino acid sequence of mDRA prompted us to investigate mDRA-associated regulatory proteins using yeast-based two-hybrid system. Two candidate clones containing mouse cDNA insert of
PAX 3 or PP2A B regulatory subunit delta isoform (PP2A Bฮด) were verified to interact with the COOH-end of mDRA. Like rat PP2A Bฮด (46), mouse PP2A Bฮด homolog mRNAs (2.4 kb) were abundantly expressed in testis and colon, whereas PAX 3 transcripts (6.4 kb) were expressed at high levels in heart. Patterns of tissue
distribution of mouse PP2A Bฮด homolog and PAX 3 were similar to those of mDRA. The functional relevance of PP2A Bฮด to mDRA as a negative regulator of tumorigenesis led us to clone the full-length of mouse PP2A Bฮด homolog cDNA. The full-length open reading frame as well as 31 nucleotides of 5'-untranslated and 565 nucleotides of 3'-untranslated were obtained using an reverse transcription-polymerase chain reaction (RT-PCR) of mouse colon cDNA. The mouse PP2A Bฮด homolog cDNA encodes a protein of 453 amino acids with a calculated Mr of 51,954 (accession number
AF366393). An amino acid alignment of mouse PP2A Bฮด reveals 96๏ผ
identity with rat PP2A Bฮด. The heterologous functional expression of this full-length PP2A Bฮด cDNA along with the core subunits of A and C in the mDRA stable transfectants will allow
us to study how PP2A holoenzyme binding to mDRA can modulate mDRA function.
Taken all together, the identification and characterization of the associating proteins with mDRA, suggestive of potential regulatory proteins can be the essential study to elucidate the regulatory mechanism of mDRA function.ope
๋ํ ๊ต์์์ ๊ณผ์ ์ ๊ฐ์ ํ์์ฑ์ ๋ํ ์ฐ๊ตฌ : ๋ํ๋ณ ํํฉ ๋ฐ ํ์๋ค์ ์ธ์๋์ ๋ํ ์กฐ์ฌ
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ผ๋ฌธ(์์ฌ)--์์ธ๋ํ๊ต ๋ํ์ :๊ต์กํ๊ณผ ์์
๊ต์ก์ ๊ณต,2002.Maste