Cloning and molecular characterization of Dashurin encoded by C20orf116, a PCI-domain containing protein.
BACKGROUND
Characterization of gene products originating from undefined open reading frames and delineation of biological functions has become the task after the human genome has been decoded.


METHODS
We cloned the human C20orf 116 and defined its transcript in liver, kidney and various brain regions by Northern analysis. Antibodies against recombinant protein used for immunofluorescence and immunoblots confirmed its expression in these tissues. With the focus on kidney, its tubular expression and presence in glomerula were shown.


RESULTS
A 28 aa long signal peptide predicted by in silico analysis is reflected by visualization of size variants of approximately 3kDa difference suggesting a signal peptidase cleavage of the proform. Cell compartment separation confirmed the presence of Dashurin in peroxisomes/mitochondria, microsomes, cytosol and nucleus. This is in line with green fluorescent protein (GFP)-Dashurin fusion protein shuttling between cytosol and nucleus. Luciferase reporter studies revealed a 2-3 fold increase of promoter activities upon over-expression. Bioinformatic analysis identified a PCI-domain at the C-terminus providing protein-protein interaction capabilities.


CONCLUSIONS
Our present findings suggest the involvement of Dashurin in gene transcription or mRNA translation.


CONCLUSIONS
Dashurin shares the PCI-domain with three multisubunit protein complexes (26S proteasome, COP9 signalosome and eIF3 translation initiation factor).
Neziri
D
D
Medical University of Vienna, Austria; Department of Internal Medicine III, Austria.
Ilhan
A
A
Maj
M
M
Majdic
O
O
Baumgartner-Parzer
S
S
Cohen
G
G
Base
W
W
Wagner
L
L
D048868:Adaptor Proteins, Signal Transducing; D006528:Carcinoma, Hepatocellular / Q000235:genetics; D045744:Cell Line, Tumor; D002874:Chromosome Mapping; D002890:Chromosomes, Human, Pair 20 / Q000235:genetics*; D003001:Cloning, Molecular / Q000379:methods; D015894:Genome, Human; D049452:Green Fluorescent Proteins / Q000235:genetics; D006801:Humans; D007668:Kidney / Q000502:physiology*; D015470:Leukemia, Myeloid, Acute / Q000235:genetics; D008113:Liver Neoplasms / Q000235:genetics; D008156:Luciferases / Q000235:genetics; D016366:Open Reading Frames; D046988:Proteasome Endopeptidase Complex / Q000235:genetics*; D014176:Protein Biosynthesis; D011506:Proteins / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000235:genetics; D014158:Transcription, Genetic; D014162:Transfection; D014178:Translocation, Genetic
D048868:Adaptor Proteins, Signal Transducing; C000622456:DDRGK1 protein, human; D011506:Proteins; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D008156:Luciferases; D046988:Proteasome Endopeptidase Complex; C061553:ATP dependent 26S protease