Wnt Signals Can Function as Positional Cues in Establishing Cell Polarity

Wnt signaling plays important roles in cell polarization in diverse organisms, and loss of cell polarity is an early event in tumorigenesis caused by mutations in Wnt pathway genes. Despite this, the precise roles of Wnt proteins in cell polarization have remained elusive. In no organism has it been shown that the asymmetric position of a Wnt signal is essential to establishing a cell’s polarity. Attempts to test this by ubiquitous expression of Wnt genes have suggested that Wnt signals might act only as permissive factors in cell polarization. Here we find, using cell manipulations and ectopic gene expression in C. elegans, that the position from which Wnt signals are presented can determine the polarity of both embryonic and postembryonic cells. Furthermore, the position from which a Wnt signal is presented can determine the polarity of Frizzled receptor localization, suggesting that the polarizing effect of Wnt is likely to be direct. These results demonstrate that Wnt proteins can function as positional cues in establishing cell polarity

Cyclin E and CDK2 Repress the Terminal Differentiation of Quiescent Cells after Asymmetric Division in C. elegans

Coordination between cell proliferation and differentiation is important in normal development and oncogenesis. These processes usually have an antagonistic relationship, in that differentiation is blocked in proliferative cells, and terminally differentiated cells do not divide. In some instances, cyclins, cyclin-dependent kinases (CDKs) and their inhibitors (CKIs) play important roles in this antagonistic regulation. However, it is unknown whether CKIs and cyclin/CDKs regulate the uncommitted state in quiescent cells where CDK activities are likely to be low. Here, we show in C. elegans that cye-1/cyclin E and cdk-2/CDK2 repress terminal differentiation in quiescent cells. In cye-1 mutants and cdk-2(RNAi) animals, after asymmetric division, certain quiescent cells adopted their sister cells' phenotype and differentiated at some frequency. In contrast, in cki-1(RNAi) animals, these cells underwent extra divisions, while, in cki-1(RNAi); cdk-2(RNAi) or cki-1(RNAi); cye-1 animals, they remained quiescent or differentiated. Therefore, in wild-type animals, CKI-1/CKI in these cells maintained quiescence by inhibiting CYE-1/CDK-2, while sufficient CYE-1/CDK-2 remained to repress the terminal differentiation. The difference between sister cells is regulated by the Wnt/MAP kinase pathway, which causes asymmetric expression of CYE-1 and CKI-1. Our results suggest that the balance between the levels of CKI and cyclin E determines three distinct cell states: terminally differentiated, quiescent and uncommitted, and proliferating

Multiple Wnts Redundantly Control Polarity Orientation in Caenorhabditis elegans Epithelial Stem Cells

During development, cell polarization is often coordinated to harmonize tissue patterning and morphogenesis. However, how extrinsic signals synchronize cell polarization is not understood. In Caenorhabditis elegans, most mitotic cells are polarized along the anterior-posterior axis and divide asymmetrically. Although this process is regulated by a Wnt-signaling pathway, Wnts functioning in cell polarity have been demonstrated in only a few cells. We analyzed how Wnts control cell polarity, using compound Wnt mutants, including animals with mutations in all five Wnt genes. We found that somatic gonadal precursor cells (SGPs) are properly polarized and oriented in quintuple Wnt mutants, suggesting Wnts are dispensable for the SGPs' polarity, which instead requires signals from the germ cells. Thus, signals from the germ cells organize the C. elegans somatic gonad. In contrast, in compound but not single Wnt mutants, most of the six seam cells, V1–V6 (which are epithelial stem cells), retain their polarization, but their polar orientation becomes random, indicating that it is redundantly regulated by multiple Wnt genes. In contrast, in animals in which the functions of three Wnt receptors (LIN-17, MOM-5, and CAM-1) are disrupted—the stem cells are not polarized and divide symmetrically—suggesting that the Wnt receptors are essential for generating polarity and that they function even in the absence of Wnts. All the seam cells except V5 were polarized properly by a single Wnt gene expressed at the cell's anterior or posterior. The ectopic expression of posteriorly expressed Wnts in an anterior region and vice versa rescued polarity defects in compound Wnt mutants, raising two possibilities: one, Wnts permissively control the orientation of polarity; or two, Wnt functions are instructive, but which orientation they specify is determined by the cells that express them. Our results provide a paradigm for understanding how cell polarity is coordinated by extrinsic signals

Transformation of quiescent cells to DTCs after their divisions in <i>cye-1</i> mutants.

<p>(A–K) Anterior is to the left; ventral is to the bottom. Merged GFP and Nomarski images. The gonad is outlined with dotted lines. The original DTC (Z1.aa) and its sister cell (Z1.ap) are marked by an arrowhead and arrow, respectively. The extra <i>lag-2::GFP</i>-positive cells produced from Z1.ap (J) or Z1.p (K) are indicated by asterisks. Scale bar, 10 µm. (A–H) Real-time analyses of <i>lag-2::GFP</i> expression in wild type (A–C), <i>cye-1(os66)</i> mutants (D–F), and <i>hs::cki-1</i> animals after heat shock (G and H) from the late L1 to early L2 stage. Each vertical set of panels represents the same animal over time. The expression about 3 hours (A and D), 5 hours (B, E and G), and 8 hours (C, F and H) after division of the Z1.a cell is shown. (I–K) <i>lag-2::GFP</i> expression in <i>cdk-2(RNAi)</i> (I) and <i>cki-1(RNAi)</i> animals (J and K).</p

Multiple functions of PBRM-1/Polybromo- and LET-526/Osa-containing chromatin remodeling complexes in C. elegans development

AbstractThe SWI/SNF-like chromatin remodeling complexes consist of two evolutionarily conserved subclasses, which are characterized by specific accessory components, the OSA/BAF250 and Polybromo proteins. These complexes regulate the expressions of distinct sets of target genes, with some overlap, and the regulatory components are thought to determine the target specificity for each complex. Here we isolated C. elegans mutants of the genes for the OSA/BAF250 homolog, LET-526, and the Polybromo homolog, PBRM-1, in a screen for the abnormal asymmetric cell division phenotype. In the asymmetric division of the T cell, both LET-526 and PBRM-1 regulated the asymmetric expression of psa-3/Meis between the T cell daughters, suggesting that the two subclasses share the same target. In the gonad, PBRM-1 regulated gonad primordium formation during embryogenesis, whereas LET-526 was required post-embryonically for distal tip cell (DTC) production from the gonad primordium, suggesting that these proteins have distinct targets for DTC development. Thus, the same cellular process is regulated by LET-526 and PBRM-1 in the asymmetric division of the T cell, but they regulate distinct cellular processes in the gonad morphogenesis. Although disruption of the core component PSA-1 or PSA-4 caused similar defects in the gonad and T cell, it also caused early embryonic arrest, which was not observed in the let-526, pbrm-1, or let-526 pbrm-1 double mutants, suggesting that some targets of SWI/SNF-like complexes do not require LET-526 or PBRM-1 for their transcription. Our results show that the target selection by SWI/SNF-like complexes during C. elegans development is intricately regulated by accessory components