TY - JOUR
T1 - Phosphorylation of Pnut in the early stages of Drosophila embryo development affects association of the septin complex with the membrane and is important for viability
AU - Akhmetova, Katarina
AU - Balasov, Maxim
AU - Svitin, Anton
AU - Chesnokova, Elena
AU - Renfrow, Matthew
AU - Chesnokov, Igor
N1 - Copyright © 2018 Akhmetova et al.
PY - 2018/1/4
Y1 - 2018/1/4
N2 - Septin proteins are polymerizing GTPases that are found in most eukaryotic species. Septins are important for cytokinesis and participate in many processes involving spatial modifications of the cell cortex. In Drosophila, septin proteins Pnut, Sep1, and Sep2 form a hexameric septin complex. Here, we found that septin protein Pnut is phosphorylated during the first 2 hr of Drosophila embryo development. To study the effect of Pnut phosphorylation in a live organism, we created a new Drosophila pnut null mutant that allows for the analysis of Pnut mutations during embryogenesis. To understand the functional significance of Pnut phosphorylation, Drosophila strains carrying nonphosphorylatable and phosphomimetic mutant pnut transgenes were established. The expression of the nonphosphorylatable Pnut protein resulted in semilethality and abnormal protein localization, whereas the expression of the phospho-mimetic mutant form of Pnut disrupted the assembly of a functional septin complex and septin filament formation in vitro. Overall, our findings indicate that the controlled phosphorylation of Pnut plays an important role in regulating septin complex functions during organism development.
AB - Septin proteins are polymerizing GTPases that are found in most eukaryotic species. Septins are important for cytokinesis and participate in many processes involving spatial modifications of the cell cortex. In Drosophila, septin proteins Pnut, Sep1, and Sep2 form a hexameric septin complex. Here, we found that septin protein Pnut is phosphorylated during the first 2 hr of Drosophila embryo development. To study the effect of Pnut phosphorylation in a live organism, we created a new Drosophila pnut null mutant that allows for the analysis of Pnut mutations during embryogenesis. To understand the functional significance of Pnut phosphorylation, Drosophila strains carrying nonphosphorylatable and phosphomimetic mutant pnut transgenes were established. The expression of the nonphosphorylatable Pnut protein resulted in semilethality and abnormal protein localization, whereas the expression of the phospho-mimetic mutant form of Pnut disrupted the assembly of a functional septin complex and septin filament formation in vitro. Overall, our findings indicate that the controlled phosphorylation of Pnut plays an important role in regulating septin complex functions during organism development.
KW - Drosophila
KW - Orc6
KW - Phosphorylation
KW - Pnut
KW - Septins
KW - Amino Acid Sequence
KW - Cell Line
KW - Drosophila melanogaster/cytology
KW - Protein Multimerization
KW - Cytoplasm/metabolism
KW - Septins/genetics
KW - Cytokinesis
KW - Macrophages/cytology
KW - Microfilament Proteins/deficiency
KW - Drosophila Proteins/deficiency
KW - Animals
KW - Gene Expression Regulation, Developmental
KW - Cell Membrane/metabolism
KW - Embryo, Nonmammalian
KW - Cytoskeleton/metabolism
KW - Protein Binding
KW - Mutation
UR - http://www.scopus.com/inward/record.url?scp=85039927269&partnerID=8YFLogxK
U2 - 10.1534/g3.117.300186
DO - 10.1534/g3.117.300186
M3 - Article
C2 - 29079679
AN - SCOPUS:85039927269
VL - 8
SP - 27
EP - 38
JO - G3: Genes, Genomes, Genetics
JF - G3: Genes, Genomes, Genetics
SN - 2160-1836
IS - 1
ER -