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Quantification and regulation of mRNAs encoding beaded filament proteins in the chick lens
M E Ireland1, D J Goebel, H Maisel
1Department of Anatomy and Cell Biology, Wayne State University School of Medicine, Detroit, MI 48201, USA.
Purpose:
To quantify the expression of beaded filament protein mRNA levels in regions of the chick lens and to examine the in vitro regulation of message and protein levels using cell culture techniques.
Methods:
RNase protection assays and Northern blotting were used to quantify beaded filament protein mRNA levels in dissected lenses. Cultured cells were assayed for mRNA with RNase protection and for protein with Western blotting and ELISA techniques after treatment with cAMP analogs.
Results:
Beaded filament protein message levels were greatly up-regulated in cortical fiber cells compared to annular pad cells. Full length messages were also detected in nuclear fiber cells. The presence of an unusual form of the CP49 message with a lamin-like insert, CP49INS, was also established. Both message and protein levels were subject to regulation in response to elevated intracellular cAMP levels.
Conclusions:
The accumulation of beaded filament protein levels during fiber cell development may be due to the increased cAMP-mediated transcription of message. The presence of CP49INS may lend new insight into mechanisms of intermediate filament assembly.