Analysis of the 3’ untranslated regions of α-tubulin and S-crystallin mRNA and the identification of CPEB in dark- and light-adapted octopus retinas

نویسندگان

  • Shannan Kelly
  • Hideki Yamamoto
  • Laura J. Robles
چکیده

PURPOSE We previously reported the differential expression and translation of mRNA and protein in dark- and light-adapted octopus retinas, which may result from cytoplasmic polyadenylation element (CPE)-dependent mRNA masking and unmasking. Here we investigate the presence of CPEs in alpha-tubulin and S-crystallin mRNA and report the identification of cytoplasmic polyadenylation element binding protein (CPEB) in light- and dark-adapted octopus retinas. METHODS 3'-RACE and sequencing were used to isolate and analyze the 3'-UTRs of alpha-tubulin and S-crystallin mRNA. Total retinal protein isolated from light- and dark-adapted octopus retinas was subjected to western blot analysis followed by CPEB antibody detection, PEP-171 inhibition of CPEB, and dephosphorylation of CPEB. RESULTS The following CPE-like sequence was detected in the 3'-UTR of isolated long S-crystallin mRNA variants: UUUAACA. No CPE or CPE-like sequences were detected in the 3'-UTRs of alpha-tubulin mRNA or of the short S-crystallin mRNA variants. Western blot analysis detected CPEB as two putative bands migrating between 60-80 kDa, while a third band migrated below 30 kDa in dark- and light-adapted retinas. CONCLUSIONS The detection of CPEB and the identification of the putative CPE-like sequences in the S-crystallin 3'-UTR suggest that CPEB may be involved in the activation of masked S-crystallin mRNA, but not in the regulation of alpha-tubulin mRNA, resulting in increased S-crystallin protein synthesis in dark-adapted octopus retinas.

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عنوان ژورنال:
  • Molecular Vision

دوره 14  شماره 

صفحات  -

تاریخ انتشار 2008