Expression of ADAMTS metalloproteinases in the retinal pigment epithelium derived cell line ARPE-19: transcriptional regulation by TNFalpha.
ADAMTS (A Disintegrin-like And Metalloprotease domain with ThromboSpondin type I motifs) are multidomain proteins with demonstrated metalloproteinase functionality and have potential roles in embryonic development, angiogenesis and cartilage degradation. We present here investigations of ADAMTS expression in an ocular cell type, ARPE-19, with a view to implicating them in retinal matrix turnover. Expression analysis was undertaken using a combination of reverse transcription polymerase chain reaction (RT-PCR) and Northern blotting experiments, which together detected the expression of mRNAs for several ADAMTS proteins, all of which have active site motifs characteristic of matrix metalloproteases (MMPs). These included ADAMTS1, ADAMTS2, ADAMTS3, ADAMTS5, ADAMTS6, ADAMTS7 and ADAMTS9. The expression of mRNA isoforms for ADAMTS7 and ADAMTS9 were also detected. Following stimulation with TNFalpha, ADAMTS1, ADAMTS6 and both ADAMTS9 transcripts expressed in ARPE-19 cells showed a potent upregulation. The expression of ADAMTS genes in ARPE-19 cells and the transcriptional stimulation of some family members by TNFalpha may implicate them in inflammatory eye disease and the compromise of retinal matrix structure, which is evident in age-related macular degeneration (ARMD) and other retinal pathologies.
Bevitt
Debra J
DJ
Department of Rheumatology, University of Newcastle Medical School, Framlington Place, NE2 4HH, Newcastle, UK.
Mohamed
Jeseem
J
Catterall
Jon B
JB
Li
Zheng
Z
Arris
Christine E
CE
Hiscott
Paul
P
Sheridan
Carl
C
Langton
Kevin P
KP
Barker
Michael D
MD
Clarke
Michael P
MP
McKie
Norman
N
D051722:ADAM Proteins; D000071118:ADAMTS7 Protein; D000071119:ADAMTS9 Protein; D015152:Blotting, Northern; D002460:Cell Line; D005786:Gene Expression Regulation; D006801:Humans; D008666:Metalloendopeptidases / Q000096:biosynthesis / Q000235:genetics*; D010857:Pigment Epithelium of Eye / Q000187:drug effects / Q000378:metabolism*; D020033:Protein Isoforms / Q000096:biosynthesis / Q000235:genetics; D012333:RNA, Messenger / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014158:Transcription, Genetic; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*
D020033:Protein Isoforms; D012333:RNA, Messenger; D014409:Tumor Necrosis Factor-alpha; D051722:ADAM Proteins; D000071118:ADAMTS7 Protein; C121534:ADAMTS7 protein, human; D000071119:ADAMTS9 Protein; C412869:ADAMTS9 protein, human; D008666:Metalloendopeptidases