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PMID: 9826689 Published · ppublish English Journal Article

Targeting cyclin-dependent kinases in Drosophila with peptide aptamers.

Kolonin MG, Finley RL

Abstract

Two-hybrid technology provides a simple way to isolate small peptide aptamers that specifically recognize and strongly bind to a protein of interest. These aptamers have the potential to dominantly interfere with specific activities of their target proteins and, therefore, could be used as in vivo inhibitors. Here we explore the ability to use peptide aptamers as in vivo inhibitors by expressing aptamers directed against cell cycle regulators in Drosophila. We expressed two peptide aptamers, each of which specifically recognizes one of the two essential cyclin-dependent kinases (Cdks), DmCdk1 and DmCdk2, in Drosophila. Expression of each Cdk aptamer during organogenesis caused adult eye defects typical of those caused by cell cycle inhibition. Co-overexpression of DmCdk1 or DmCdk2 resulted in suppression of the eye phenotypes, indicating that each aptamer interacts with a Cdk target in vivo and suggesting that these peptides disrupt normal eye development by inhibiting Cdk function. Moreover, the specificity of each aptamer for one of the two Cdks as determined in two-hybrid assays was retained in Drosophila. Combined, our results demonstrate that peptide aptamers generated by yeast two-hybrid methods can serve as inhibitory reagents to target specific proteins in vivo.

MeSH Terms
Animals Animals, Genetically Modified CDC2 Protein Kinase/biosynthesis,genetics CDC2-CDC28 Kinases Chromosome Mapping Crosses, Genetic Cyclin-Dependent Kinase 2 Cyclin-Dependent Kinases/biosynthesis,genetics Drosophila/enzymology,genetics,growth & development Drosophila Proteins Eye/anatomy & histology Eye Abnormalities/chemically induced,genetics Female Homozygote Male Peptides/pharmacology Protein Serine-Threonine Kinases/biosynthesis,genetics X Chromosome
Chemicals
Drosophila Proteins Peptides Protein Serine-Threonine Kinases CDC2 Protein Kinase CDC2-CDC28 Kinases Cyclin-Dependent Kinase 2 Cyclin-Dependent Kinases cdk2 protein, Drosophila
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Kolonin M G
Center for Molecular Medicine and Genetics, Wayne State University School of Medicine, 540 East Canfield Avenue, Detroit, MI 48201, USA.
Finley R L
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
Published
1998-11-24
Pages
14266-71
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC24362
Subset
IM
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