OWL1: an Arabidopsis J-domain protein involved in perception of very low light fluences.

To sense ambient light conditions in order to optimize their growth and development, plants employ a battery of photoreceptors responsive to light quality and quantity. Essential for the sensing of red and far-red (FR) light is the phytochrome family of photoreceptors. Among them, phytochrome A is special because it mediates responses to different light conditions, including both very low fluences (very low fluence response [VLFR]) and high irradiances (high irradiance response [HIR]). In contrast with the FR-HIR signaling pathway, in which several intermediates of the signaling pathway have been identified, specific components of the VLFR pathway remain unknown. Here, we describe owl1 (for orientation under very low fluences of light), a mutant that is specific for the VLFR, suggesting that VLFR and HIR pathways are genetically distinct, although some common mechanisms can be observed. OWL1 codes for a ubiquitous J-domain protein essential for germination, cotyledon opening, hypocotyl elongation, and deviation of the direction of hypocotyl growth from the vertical under very low light conditions. Additionally, we observed a flowering phenotype suggesting a role for the VLFR during the whole life cycle of a plant. OWL1 interacts with the basic helix-loop-helix HFR1 (LONG HYPOCOTYL IN FAR-RED) transcription factor, previously characterized as a component of the FR-HIR pathway. Both proteins are involved in the agravitropic response under FR light. We propose a central function of OWL1 in the VLFR pathway, which is essential for plant survival under unfavorable light conditions.

Cell cycle-dependent roles for the FCH-domain protein Cdc15p in formation of the actomyosin ring in Schizosaccharomyces pombe.

Cell division in the fission yeast Schizosaccharomyces pombe requires the formation and constriction of an actomyosin ring at the division site. The actomyosin ring is assembled in metaphase and anaphase A, is maintained throughout mitosis, and constricts after completion of anaphase. Maintenance of the actomyosin ring during late stages of mitosis depends on the septation initiation network (SIN), a signaling cascade that also regulates the deposition of the division septum. However, SIN is not active in metaphase and is not required for the initial assembly of the actomyosin ring early in mitosis. The FER/CIP4-homology (FCH) domain protein Cdc15p is a component of the actomyosin ring. Mutations in cdc15 lead to failure in cytokinesis and result in the formation of elongated, multinucleate cells without a division septum. Here we present evidence that the requirement of Cdc15p for actomyosin ring formation is dependent on the stage of mitosis. Although cdc15 mutants are competent to assemble actomyosin rings in metaphase, they are unable to maintain actomyosin rings late in mitosis when SIN is active. In the absence of functional Cdc15p, ring formation upon metaphase arrest depends on the anillin-like Mid1p. Interestingly, when cytokinesis is delayed due to perturbations to the division machinery, Cdc15p is maintained in a hypophosphorylated form. The dephosphorylation of Cdc15p, which occurs transiently in unperturbed cytokinesis, is partially dependent on the phosphatase Clp1p/Flp1p. This suggests a mechanism where both SIN and Clp1p/Flp1p contribute to maintenance of the actomyosin ring in late mitosis through Cdc15p, possibly by regulating its phosphorylation status.

Yeast lipid rafts?--an emerging view.

In various eukaryotes, sterol-rich membrane domains have been proposed to play an important role in polarization and compartmentalization of the plasma membrane. Several studies have reported the cellular distribution of sterols in genetically tractable yeast species and the identification of molecules that might regulate the localization of sterol-rich membrane domains. Here, we attempt to synthesize our understanding of the function and organization of these domains from the study of fungi and identify some outstanding issues.

The novel fission yeast protein Pal1p interacts with Hip1-related Sla2p/End4p and is involved in cellular morphogenesis.

The establishment and maintenance of characteristic cellular morphologies is a fundamental property of all cells. Here we describe Schizosaccharomyces pombe Pal1p, a protein important for maintenance of cylindrical cellular morphology. Pal1p is a novel membrane-associated protein that localizes to the growing tips of interphase cells and to the division site in cells undergoing cytokinesis in an F-actin- and microtubule-independent manner. Cells deleted for pal1 display morphological defects, characterized by the occurrence of spherical and pear-shaped cells with an abnormal cell wall. Pal1p physically interacts and displays overlapping localization with the Huntingtin-interacting-protein (Hip1)-related protein Sla2p/End4p, which is also required for establishment of cylindrical cellular morphology. Sla2p is important for efficient localization of Pal1p to the sites of polarized growth and appears to function upstream of Pal1p. Interestingly, spherical pal1Delta mutants polarize to establish a pearlike morphology before mitosis in a manner dependent on the kelch-repeat protein Tea1p and the cell cycle inhibitory kinase Wee1p. Thus, overlapping mechanisms involving Pal1p, Tea1p, and Sla2p contribute to the establishment of cylindrical cellular morphology, which is important for proper spatial regulation of cytokinesis.

Sterol-rich plasma membrane domains in the fission yeast Schizosaccharomyces pombe.

Sterol-rich membrane domains exist in unicellular and multicellular eukaryotes. They are thought to provide a structural framework for interactions among a subset of proteins by selectively incorporating some proteins while excluding others. Although most studies have focused on the biophysical and biochemical properties of sterol-rich membrane domains and incorporated proteins, relatively little is known about their intracellular distribution. Using a cytological approach we show here that in the fission yeast Schizosaccharomyces pombe, sterols are enriched in the plasma membrane at the growing cell tips and at the site of cytokinesis. The distribution of sterols is regulated in a cell-cycle-dependent manner and requires a functional secretory pathway. By manipulating the integrity of sterol-rich membrane domains using sterol sequestering agents and genetic means, we show that these domains are important for multiple processes regulating cytokinesis. In these cells, defects in proper maintenance of the actomyosin ring and/or its attachment to the overlying plasma membrane were observed. Furthermore, the stability of a plasma membrane protein that colocalises with sterol-rich membrane domains was compromised. Taken together, our studies establish S. pombe as a genetically tractable model organism in which to study the role(s) of sterol-rich membrane domains in cell polarity and cytokinesis.