A chemical-genetics approach to study the role of atypical Protein Kinase C in Drosophila

• Published in: Development (Cambridge) Vol. 146; no. 2
• Main Authors: Hannaford, Matthew, Loyer, Nicolas, Tonelli, Francesca, Zoltner, Martin, Januschke, Jens
• Format: Journal Article
• Language: English
• Published: England The Company of Biologists Ltd 15.01.2019
• Subjects: Alleles; Animals; Cell Division; Cell Polarity; Drosophila melanogaster - embryology; Drosophila melanogaster - enzymology; Drosophila melanogaster - genetics; Drosophila Proteins - antagonists & inhibitors; Drosophila Proteins - genetics; Drosophila Proteins - metabolism; Embryo, Nonmammalian - cytology; Embryo, Nonmammalian - metabolism; Larva - cytology; Larva - metabolism; Loss of Function Mutation - genetics; Neurons - metabolism; Protein Kinase C - antagonists & inhibitors; Protein Kinase C - genetics; Protein Kinase C - metabolism; RNA Interference; Techniques and Resources; Chemical genetics; Neuroblasts; Drosophila; atypical Protein Kinase C; Asymmetric cell division
• ISSN: 0950-1991; 1477-9129
• DOI: 10.1242/dev.170589

Studying the function of proteins using genetics in cycling cells is complicated by the fact that there is often a delay between gene inactivation and the time point of phenotypic analysis. This is particularly true when studying kinases that have pleiotropic functions and multiple substrates. neuroblasts (NBs) are rapidly dividing stem cells and an important model system for the study of cell polarity. Mutations in multiple kinases cause NB polarity defects, but their precise functions at particular time points in the cell cycle are unknown. Here, we use chemical genetics and report the generation of an analogue-sensitive allele of atypical Protein Kinase C (aPKC). We demonstrate that the resulting mutant aPKC kinase can be specifically inhibited and Acute inhibition of aPKC during NB polarity establishment abolishes asymmetric localization of Miranda, whereas its inhibition during NB polarity maintenance does not in the time frame of normal mitosis. However, aPKC helps to sharpen the pattern of Miranda, by keeping it off the apical and lateral cortex after nuclear envelope breakdown.