Within the Arabidopsis thaliana family of 21 heat stress transcription factors (Hsfs), HsfA9 is exclusively expressed in late stages of seed development. Here, we present evidence that developmental expression of HsfA9 is regulated by the seed-specific transcription factor ABSCISIC ACID–INSENSITIVE3 (ABI3). Intriguingly, ABI3 knockout lines lack detectable levels of HsfA9 transcript and protein, and further ectopic expression of ABI3 conferred the ability to accumulate HsfA9 in response to abscisic acid in transgenic plantlets. Consequently, the most abundant heat stress proteins (Hsps) in seeds (Hsp17.4-CI, Hsp17.7-CII, and Hsp101) were not detectable in the ABI3 knockout lines, but their expression could be detected in plants ectopically expressing HsfA9 in vegetative tissues. Furthermore, this seed-specific transcription factor cascade was reconstructed in transient β-glucuronidase reporter assays in mesophyll protoplasts by showing that ABI3 could activate the HsfA9 promoter, whereas HsfA9 in turn was shown to be a potent activator on the promoters of Hsp genes. Thus, our study establishes a genetic framework in which HsfA9 operates as a specialized Hsf for the developmental expression of Hsp genes during seed maturation.

Correct spindle positioning is fundamental for proper cell division during development and in stem cell lineages. Dynein and an evolutionarily conserved ternary complex (nuclear mitotic apparatus protein [NuMA]–LGN–Gα in human cells and LIN-5–GPR-1/2–Gα in Caenorhabditis elegans) are required for correct spindle positioning, but their relationship remains incompletely understood. By analyzing fixed specimens and conducting live-imaging experiments, we uncovered that appropriate levels of ternary complex components are critical for dynein-dependent spindle positioning in HeLa cells and C. elegans embryos. Moreover, using mutant versions of Gα in both systems, we established that dynein acts at the membrane to direct spindle positioning. Importantly, we identified a region within NuMA that mediates association with dynein. By using this region to target dynein to the plasma membrane, we demonstrated that the mere presence of dynein at that location is sufficient to direct spindle positioning in HeLa cells. Overall, we propose a model in which the ternary complex serves to anchor dynein at the plasma membrane to ensure correct spindle positioning.