Search for: All records

ABSTRACT Of the three types of cytoskeleton known in animals—actin, microtubules, and intermediate filaments—only actin and microtubules exist in plants. Both play important roles in cellular shaping, organelle movement, organization of the endomembrane system, and cell signaling. An emerging, but often overlooked role of the plant cytoskeleton is its dynamic and mutually influential interaction with the nucleus. Here, we summarize recent advances in understanding the role of the cytoskeleton in plant nuclear movement in different biological contexts, a role for nuclear envelope‐associated proteins in reorganizing the actin and microtubule cytoskeleton, and the molecular nature of the nucleus‐cytoskeleton interface and specific proteins contributing to it. In animals, the nucleoskeleton consists of the nuclear lamina, an intermediate‐filament meshwork underlying the nuclear envelope. Plants have evolved an equivalent of this structure, built by different types of proteins. Here, we highlight recent advances in understanding its filamentous organization, newly discovered protein interactions connecting it to nuclear pores, and exciting new evidence that—just like the animal lamina—the plant lamina is involved in chromatin reorganization and epigenetic changes. Together, these new developments create new opportunities toward a deeper understanding of this important regulatory connection between the cytoskeleton and the cell's largest organelle. 

ABSTRACT Of the 61 kinesins annotated inArabidopsis thaliana, many are still without assigned function. Here, we have screened an insertional mutant library of Arabidopsis pollen‐expressed kinesins for fertility defects. Insertional mutants for three kinesins showed a significant reduction in seed set. Among them, we focused on the sole kinesin‐4 expressed in pollen (kinesin‐4C, here Pollen‐Expressed Kinesin 14, PEK14). We show a seed‐set defect in the three independent allelespek14‐1, pek14‐2, and pek14‐3. This defect is male‐derived and is equally distributed throughout the silique. Maturepek14‐1anthers contain about 10% inviable pollen grains.pek14‐1pollen tubes grow 20% more slowly and show reduced pollen tube bending. Analysis of the male germ unit (MGU), as it travels through the pollen tube, demonstrates an aberrant organization of thepek14‐1MGU in 30% of pollen tubes and an increase in the distance of the MGU to the tip by 24%. Expression of GFP‐tagged PEK14 successfully complemented the observed seed set defect, as well as the growth rate, bending, and MGU organization defects observed inpek14‐1. In pollen, PEK14‐GFP is located diffusely at the pollen tube tip. PEK14‐GFP is also expressed in the root meristematic zone and is located at the mid‐zone of the phragmoplast, but no apparent root growth phenotype was observed, likely due to redundancy in this organ. 

Key messageThe Arabidopsis KASH protein SINE3 is involved in male and female gametophyte development, likely affecting the first post-meiotic mitosis in both cases, and is required for full seed set. AbstractLinker of nucleoskeleton and cytoskeleton (LINC) complexes are protein complexes spanning the inner and outer membranes of the nuclear envelope (NE) and are key players in nuclear movement and positioning. Through their roles in nuclear movement and cytoskeletal reorganization, plant LINC complexes affect processes as diverse as pollen tube rupture and stomatal development and function. KASH proteins are the outer nuclear membrane component of the LINC complex, with conserved C-termini but divergent N-terminal cytoplasmic domains. Of the known Arabidopsis KASH proteins, SUN-INTERACTING NUCLEAR ENVELOPE PROTEIN 3 (SINE3) has not been functionally characterized. Here, we show that SINE3 is expressed at all stages of male and female gametophyte development. It is located at the NE in male and female gametophytes. Loss of SINE3 results in a female-derived seed set defect, withsine3mutant ovules arresting at stage FG1. Pollen viability is also significantly reduced, with microspores arresting prior to pollen mitosis I. In addition, sine3mutants have a minor male meiosis defect, with some tetrads containing more than four spores. Together, these results demonstrate that the KASH protein SINE3 plays a crucial role in male and female gametophyte development, likely affecting the first post-meiotic nuclear division in both cases. 

Abstract Linker of nucleoskeleton and cytoskeleton (LINC) complexes consist of outer nuclear membrane KASH proteins, interacting in the nuclear envelope lumen with inner nuclear membrane SUN proteins and connecting the nucleus and cytoskeleton. The paralogous Arabidopsis KASH proteins SINE1 and SINE2 function during stomatal dynamics induced by light–dark transitions and abscisic acid (ABA), which requires F-actin reorganization. SINE2 influences actin depolymerization and SINE1 actin repolymerization. The actin-related protein 2/3 (ARP2/3) complex, an actin nucleator, and the plant actin-bundling and -stabilizing factor SCAB1 are involved in stomatal aperture control. Here, we have tested the genetic interaction of SINE1 and SINE2 with SCAB1 and the ARP2/3 complex. We show that SINE1 and the ARP2/3 complex function in the same pathway during ABA-induced stomatal closure, while SINE2 and the ARP2/3 complex play opposing roles. The actin repolymerization defect observed in sine1-1 is partially rescued in scab1-2 sine1-1, while SINE2 is epistatic to SCAB1. In addition, SINE1 and ARP2/3 act synergistically in lateral root development. The absence of SINE2 renders trichome development independent of the ARP2/3 complex. Together, these data reveal complex and differential interactions of the two KASH proteins with the actin-remodeling apparatus and add evidence to the proposed differential role of SINE1 and SINE2 in actin dynamics. 

The linker of nucleoskeleton and cytoskeleton (LINC) complex is a protein complex spanning the inner and outer membranes of the nuclear envelope. Outer nuclear membrane KASH proteins interact in the nuclear envelope lumen with inner nuclear membrane SUN proteins. The paralogous Arabidopsis KASH proteins SINE1 and SINE2 function during stomatal dynamics induced by light–dark transitions and ABA. Previous studies have shown F-actin organization, cytoplasmic calcium (Ca 2+ ) oscillations, and vacuolar morphology changes are involved in ABA-induced stomatal closure. Here, we show that SINE1 and SINE2 are both required for actin pattern changes during ABA-induced stomatal closure, but influence different, temporally distinguishable steps. External Ca 2+ partially overrides the mutant defects. ABA-induced cytoplasmic Ca 2+ oscillations are diminished in sine2-1 but not sine1-1 , and this defect can be rescued by both exogenous Ca 2+ and F-actin depolymerization. We show first evidence for nuclear Ca 2+ oscillations during ABA-induced stomatal closure, which are disrupted in sine2-1 . Vacuolar fragmentation is impaired in both mutants and is partially rescued by F-actin depolymerization. Together, these data indicate distinct roles for SINE1 and SINE2 upstream of this network of players involved in ABA-based stomatal closure, suggesting a role for the nuclear surface in guard cell ABA signaling.