The Scaffold Proteins Paxillin B and alpha -Actinin Regulate Septation in Aspergillus nidulans via Control of Actin Ring Contraction.

Cytokinesis, as the final step of cell division, plays an important role in fungal growth and proliferation. In the filamentous fungus Aspergillus nidulans, defective cytokinesis is able to induce abnormal multinuclear or non-nucleated cells and then result in reduced hyphal growth and abolished sporulation. Previous studies have reported that a conserved contractile actin ring (CAR) protein complex and the septation-initiation network (SIN) signaling kinase cascade are required for cytokinesis and septation; however, little is known about the role (s) of scaffold proteins are involved in these two important cellular processes. In this study, we show that a septum-localized scaffold protein paxillin B (PaxB) is essential for cytokinesis/septation in A. nidulans. The septation defects observed in a paxB deletion strain resemble those caused by the absence of another identified scaffold protein, alpha-actinin (AcnA). Deletion of alpha-actinin (AcnA) lead to undetectable PaxB at the septation site, whereas deletion of paxB does not affect the localization of alpha-actinin at septa. However, deletion of either alpha-actinin (acnA) or paxB causes the actin ring to disappear at septation sites during cytokinesis. Notably, overexpression of alpha-actinin acnA partially rescues the septum defects of the paxB mutant but not vice versa, suggesting AcnA may play a dominant role over that of PaxB for cytokinesis and septation. In addition, PaxB and alpha-actinin affect the septal dynamic localization of MobA, a conserved component of the SIN pathway, suggesting they may affect the SIN protein complex function at septa. Protein pull-down assays combined with liquid chromatography-mass spectrometry (LC-MS) identification indicate that alpha-actinin AcnA and PaxB likely do not directly interact but presumably belong to an actin cytoskeleton protein network that are required for the assembly and contraction of the CAR. Taken together, findings in this study provide novel insights into the roles of conserved scaffold proteins during fungal septation in A. nidulans.