Arabidopsis actin-depolymerizing factor7 severs actin filaments and regulates actin cable turnover to promote normal pollen tube growth.
Actin filaments are often arranged into higher-order structures, such as the longitudinal actin cables that generate the reverse fountain cytoplasmic streaming pattern present in pollen tubes. While several actin binding proteins have been implicated in the generation of these cables, the mechanisms that regulate their dynamic turnover remain largely unknown. Here, we show that Arabidopsis thaliana actin-depolymerizing factor7 (ADF7) is required for turnover of longitudinal actin cables. In vitro biochemical analyses revealed that ADF7 is a typical ADF that prefers ADP-G-actin over ATP-G-actin. ADF7 inhibits nucleotide exchange on actin and severs filaments, but its filament severing and depolymerizing activities are less potent than those of the vegetative ADF1. ADF7 primarily decorates longitudinal actin cables in the shanks of pollen tubes. Consistent with this localization pattern, the severing frequency and depolymerization rate of filaments significantly decreased, while their maximum lifetime significantly increased, in adf7 pollen tube shanks. Furthermore, an ADF7-enhanced green fluorescent protein fusion with defective severing activity but normal G-actin binding activity could not complement adf7, providing compelling evidence that the severing activity of ADF7 is vital for its in vivo functions. These observations suggest that ADF7 evolved to promote turnover of longitudinal actin cables by severing actin filaments in pollen tubes.
Key Laboratory of Plant Molecular Physiology, Institute of Botany, Chinese Academy of Sciences, Beijing 100093, China.
Yiyan
Y
Zheng
Yurong
Y
Xie
Yuxiang
Y
Jiang
Xiaolu
X
Qu
Shanjin
S
Huang
D008841:Actin Cytoskeleton / Q000235:genetics / Q000378:metabolism*; D051339:Actin Depolymerizing Factors / Q000235:genetics* / Q000378:metabolism; D017360:Arabidopsis / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism / Q000648:ultrastructure; D029681:Arabidopsis Proteins / Q000235:genetics* / Q000378:metabolism; D019086:Bridged Bicyclo Compounds, Heterocyclic / Q000494:pharmacology; D003595:Cytoplasmic Streaming / Q000235:genetics; D018506:Gene Expression Regulation, Plant; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins; D008958:Models, Molecular; D008969:Molecular Sequence Data; D053205:Pollen Tube / Q000235:genetics / Q000254:growth & development / Q000378:metabolism; D011485:Protein Binding; D011993:Recombinant Fusion Proteins; D016415:Sequence Alignment; D053778:Thiazolidines / Q000494:pharmacology
C578063:ADF7 protein, Arabidopsis; D051339:Actin Depolymerizing Factors; D029681:Arabidopsis Proteins; D019086:Bridged Bicyclo Compounds, Heterocyclic; D011993:Recombinant Fusion Proteins; D053778:Thiazolidines; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; C037068:latrunculin B