CDNA analysis of the 49 kda Lens fiber cell cytoskeletal protein: A new, lens-specific member of the intermediate filament family?

John F. Hess, Jodi T. Casselman, Paul G FitzGerald

Research output: Contribution to journalArticle

51 Citations (Scopus)

Abstract

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.

Original languageEnglish (US)
Pages (from-to)77-88
Number of pages12
JournalCurrent Eye Research
Volume12
Issue number1
DOIs
StatePublished - 1993

Fingerprint

Cytoskeletal Proteins
Intermediate Filaments
Staphylococcal Protein A
Lenses
Reading Frames
Peptides
Proteins
Nucleic Acid Databases
Keratins
Cytoskeleton
Gene Library
DNA Sequence Analysis
Open Reading Frames
Organism Cloning
Polyacrylamide Gel Electrophoresis
Complementary DNA
Clone Cells
Molecular Weight
Immunohistochemistry
Databases

ASJC Scopus subject areas

  • Ophthalmology
  • Sensory Systems
  • Cellular and Molecular Neuroscience

Cite this

CDNA analysis of the 49 kda Lens fiber cell cytoskeletal protein : A new, lens-specific member of the intermediate filament family? / Hess, John F.; Casselman, Jodi T.; FitzGerald, Paul G.

In: Current Eye Research, Vol. 12, No. 1, 1993, p. 77-88.

Research output: Contribution to journalArticle

@article{c023f1893d0b4a7ab589ab4dced2bf98,
title = "CDNA analysis of the 49 kda Lens fiber cell cytoskeletal protein: A new, lens-specific member of the intermediate filament family?",
abstract = "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.",
author = "Hess, {John F.} and Casselman, {Jodi T.} and FitzGerald, {Paul G}",
year = "1993",
doi = "10.3109/02713689308999499",
language = "English (US)",
volume = "12",
pages = "77--88",
journal = "Current Eye Research",
issn = "0271-3683",
publisher = "Informa Healthcare",
number = "1",

}

TY - JOUR

T1 - CDNA analysis of the 49 kda Lens fiber cell cytoskeletal protein

T2 - A new, lens-specific member of the intermediate filament family?

AU - Hess, John F.

AU - Casselman, Jodi T.

AU - FitzGerald, Paul G

PY - 1993

Y1 - 1993

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=0027463042&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0027463042&partnerID=8YFLogxK

U2 - 10.3109/02713689308999499

DO - 10.3109/02713689308999499

M3 - Article

C2 - 7679620

AN - SCOPUS:0027463042

VL - 12

SP - 77

EP - 88

JO - Current Eye Research

JF - Current Eye Research

SN - 0271-3683

IS - 1

ER -