Targeting cyclin-dependent kinases in Drosophila with peptide aptamers.
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.
Kolonin
M G
MG
Center for Molecular Medicine and Genetics, Wayne State University School of Medicine, 540 East Canfield Avenue, Detroit, MI 48201, USA.
Finley
R L
RL
D000818:Animals; D030801:Animals, Genetically Modified; D016203:CDC2 Protein Kinase / Q000096:biosynthesis / Q000235:genetics*; D042846:CDC2-CDC28 Kinases; D002874:Chromosome Mapping; D003433:Crosses, Genetic; D051357:Cyclin-Dependent Kinase 2; D018844:Cyclin-Dependent Kinases / Q000096:biosynthesis / Q000235:genetics*; D004330:Drosophila / Q000201:enzymology / Q000235:genetics* / Q000254:growth & development; D029721:Drosophila Proteins; D005123:Eye / Q000033:anatomy & histology; D005124:Eye Abnormalities / Q000139:chemically induced / Q000235:genetics; D005260:Female; D006720:Homozygote; D008297:Male; D010455:Peptides / Q000494:pharmacology; D017346:Protein Serine-Threonine Kinases / Q000096:biosynthesis / Q000235:genetics*; D014960:X Chromosome
D029721:Drosophila Proteins; D010455:Peptides; D017346:Protein Serine-Threonine Kinases; D016203:CDC2 Protein Kinase; D042846:CDC2-CDC28 Kinases; D051357:Cyclin-Dependent Kinase 2; D018844:Cyclin-Dependent Kinases; C495887:cdk2 protein, Drosophila