Abstract
Purpose. Evidence linking CP 49 md CP 115 to the Beaded Filament (BF) as well as evidence for the relatedness of these two proteins to the Intermediate Filament (IF) family has been previously presented. We began experiments to determine if the yeast two hybrid system could be used to study the assembly/interaction between CP 49 and CP 115. Methods. cDNA sequences for bovine CP 115, mouse CP 49 and human vimentin were manipulated using standard molecular biology techniques and cloned into the appropriate screening plasmids. DNA constructs containing various combinations of these cDNA sequences were introduced into yeast and activity of the beta-Galactosidase reporter monitored. Results. Using human vimentin as a positive control, we have shown that the two hybrid system is useful for the study of Intermediate Filament assembly. In addition, our data show that a specific interaction occurs between CP 115 and CP 49, providing additional experimental tools to study CP 115 and CP 49 assembly. Conclusions The yeast two hybrid system provides functional evidence for an initial IF like assembly process (heteromer formation) between the lens specific Beaded Filament proteins CP 49 and CP115.
| Original language | English (US) |
|---|---|
| Journal | Investigative Ophthalmology and Visual Science |
| Volume | 37 |
| Issue number | 3 |
| State | Published - Feb 15 1996 |
Fingerprint
ASJC Scopus subject areas
- Ophthalmology
Cite this
Interaction between beaded filament proteins CP 49 and CP 115 revealed by the yeast 2 hybrid system. / Hess, J. F.; FitzGerald, Paul G.
In: Investigative Ophthalmology and Visual Science, Vol. 37, No. 3, 15.02.1996.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Interaction between beaded filament proteins CP 49 and CP 115 revealed by the yeast 2 hybrid system
AU - Hess, J. F.
AU - FitzGerald, Paul G
PY - 1996/2/15
Y1 - 1996/2/15
N2 - Purpose. Evidence linking CP 49 md CP 115 to the Beaded Filament (BF) as well as evidence for the relatedness of these two proteins to the Intermediate Filament (IF) family has been previously presented. We began experiments to determine if the yeast two hybrid system could be used to study the assembly/interaction between CP 49 and CP 115. Methods. cDNA sequences for bovine CP 115, mouse CP 49 and human vimentin were manipulated using standard molecular biology techniques and cloned into the appropriate screening plasmids. DNA constructs containing various combinations of these cDNA sequences were introduced into yeast and activity of the beta-Galactosidase reporter monitored. Results. Using human vimentin as a positive control, we have shown that the two hybrid system is useful for the study of Intermediate Filament assembly. In addition, our data show that a specific interaction occurs between CP 115 and CP 49, providing additional experimental tools to study CP 115 and CP 49 assembly. Conclusions The yeast two hybrid system provides functional evidence for an initial IF like assembly process (heteromer formation) between the lens specific Beaded Filament proteins CP 49 and CP115.
AB - Purpose. Evidence linking CP 49 md CP 115 to the Beaded Filament (BF) as well as evidence for the relatedness of these two proteins to the Intermediate Filament (IF) family has been previously presented. We began experiments to determine if the yeast two hybrid system could be used to study the assembly/interaction between CP 49 and CP 115. Methods. cDNA sequences for bovine CP 115, mouse CP 49 and human vimentin were manipulated using standard molecular biology techniques and cloned into the appropriate screening plasmids. DNA constructs containing various combinations of these cDNA sequences were introduced into yeast and activity of the beta-Galactosidase reporter monitored. Results. Using human vimentin as a positive control, we have shown that the two hybrid system is useful for the study of Intermediate Filament assembly. In addition, our data show that a specific interaction occurs between CP 115 and CP 49, providing additional experimental tools to study CP 115 and CP 49 assembly. Conclusions The yeast two hybrid system provides functional evidence for an initial IF like assembly process (heteromer formation) between the lens specific Beaded Filament proteins CP 49 and CP115.
UR - http://www.scopus.com/inward/record.url?scp=33750181589&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=33750181589&partnerID=8YFLogxK
M3 - Article
AN - SCOPUS:33750181589
VL - 37
JO - Investigative Ophthalmology and Visual Science
JF - Investigative Ophthalmology and Visual Science
SN - 0146-0404
IS - 3
ER -