cDNA analysis of the 49 kDa lens fiber cell cytoskeletal protein: a new, lens-specific member of the intermediate filament family?
Two proteins, with molecular weights of 49 (CP49) and 115 kDa (CP115) as judged by SDS PAGE, have been shown by immunocytochemistry to be components of the beaded filament, a cytoskeletal structure thus far demonstrated only in the lens fiber cell. We have used antibodies reactive with CP49 to screen a mouse lens cDNA expression library. An immunoreactive clone with an approximately 1.0 kb insert was identified and purified. DNA sequence analysis shows the presence of an open reading frame that extends from the upstream cloning site for 660 bases. Contained within this reading frame are 2 peptide sequences nearly identical to 2 peptide sequences obtained from purified bovine CP49. Northern analysis revealed that the mRNA for the CP49 is not detectable in mouse brain, muscle, lung, liver, or heart. A search of the Genbank database showed that the partial cDNA sequence for the murine CP49 is unique, but that this partial sequence shows a strong similarity to multiple members of the intermediate filament family, with greatest similarity to type I acidic cytokeratins. The data presented here suggests that the CP49 is related to, and possibly represents a new member of the intermediate filament family. These data, in concert with previously published work, suggests 1) the CP49 and CP115, which have been localized to the beaded filament, are related to the family of IF proteins, and 2) these two proteins comprise a cytoskeletal structure which is structurally distinct from classical 8-11 nm intermediate filaments, thus possibly comprising a structurally novel form of intermediate filament.
Hess
J F
JF
Department of Cell Biology and Human Anatomy, School of Medicine, University of California, Davis 95616.
Casselman
J T
JT
FitzGerald
P G
PG
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D002417:Cattle; D003459:Crystallins / Q000032:analysis* / Q000235:genetics / Q000302:isolation & purification; D004247:DNA / Q000032:analysis*; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D005786:Gene Expression Regulation; D015723:Gene Library; D007381:Intermediate Filament Proteins / Q000032:analysis* / Q000235:genetics; D007382:Intermediate Filaments / Q000737:chemistry*; D007633:Keratins / Q000235:genetics; D007908:Lens, Crystalline / Q000737:chemistry*; D051379:Mice; D008969:Molecular Sequence Data; D008970:Molecular Weight; D017386:Sequence Homology, Amino Acid
D003459:Crystallins; D007381:Intermediate Filament Proteins; D007633:Keratins; D004247:DNA