Unconventional features of C9ORF72 expanded repeat in amyotrophic lateral sclerosis and frontotemporal lobar degeneration.

Amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration (FTLD) are devastating neurodegenerative diseases that form two ends of a complex disease spectrum. Aggregation of RNA binding proteins is one of the hallmark pathologic features of ALS and FTDL and suggests perturbance of the RNA metabolism in their etiology. Recent identification of the disease-associated expansions of the intronic hexanucleotide repeat GGGGCC in the C9ORF72 gene further substantiates the case for RNA involvement. The expanded repeat, which has turned out to be the single most common genetic cause of ALS and FTLD, may enable the formation of complex DNA and RNA structures, changes in RNA transcription, and processing and formation of toxic RNA foci, which may sequester and inactivate RNA binding proteins. Additionally, the transcribed expanded repeat can undergo repeat-associated non-ATG-initiated translation resulting in accumulation of a series of dipeptide repeat proteins. Understanding the basis of the proposed mechanisms and shared pathways, as well as interactions with known key proteins such as TAR DNA-binding protein (TDP-43) are needed to clarify the pathology of ALS and/or FTLD, and make possible steps toward therapy development.

The Papaver self-incompatibility pollen S-determinant, PrpS, functions in Arabidopsis thaliana.

Many angiosperms use specific interactions between pollen and pistil proteins as "self" recognition and/or rejection mechanisms to prevent self-fertilization. Self-incompatibility (SI) is encoded by a multiallelic S locus, comprising pollen and pistil S-determinants. In Papaver rhoeas, cognate pistil and pollen S-determinants, PrpS, a pollen-expressed transmembrane protein, and PrsS, a pistil-expressed secreted protein, interact to trigger a Ca(2+)-dependent signaling network, resulting in inhibition of pollen tube growth, cytoskeletal alterations, and programmed cell death (PCD) in incompatible pollen. We introduced the PrpS gene into Arabidopsis thaliana, a self-compatible model plant. Exposing transgenic A. thaliana pollen to recombinant Papaver PrsS protein triggered remarkably similar responses to those observed in incompatible Papaver pollen: S-specific inhibition and hallmark features of Papaver SI. Our findings demonstrate that Papaver PrpS is functional in a species with no SI system that diverged ~140 million years ago. This suggests that the Papaver SI system uses cellular targets that are, perhaps, common to all eudicots and that endogenous signaling components can be recruited to elicit a response that most likely never operated in this species. This will be of interest to biologists interested in the evolution of signaling networks in higher plants.

The pollen S-determinant in Papaver: comparisons with known plant receptors and protein ligand partners.

Cell-cell communication is vital to multicellular organisms and much of it is controlled by the interactions of secreted protein ligands (or other molecules) with cell surface receptors. In plants, receptor-ligand interactions are known to control phenomena as diverse as floral abscission, shoot apical meristem maintenance, wound response, and self-incompatibility (SI). SI, in which 'self' (incompatible) pollen is rejected, is a classic cell-cell recognition system. Genetic control of SI is maintained by an S-locus, in which male (pollen) and female (pistil) S-determinants are encoded. In Papaver rhoeas, PrsS proteins encoded by the pistil S-determinant interact with incompatible pollen to effect inhibition of pollen growth via a Ca(2+)-dependent signalling network, resulting in programmed cell death of 'self' pollen. Recent studies are described here that identified and characterized the pollen S-determinant of SI in P. rhoeas. Cloning of three alleles of a highly polymorphic pollen-expressed gene, PrpS, which is linked to pistil-expressed PrsS revealed that PrpS encodes a novel approximately 20 kDa transmembrane protein. Use of antisense oligodeoxynucleotides provided data showing that PrpS functions in SI and is the pollen S-determinant. Identification of PrpS represents a milestone in the SI field. The nature of PrpS suggests that it belongs to a novel class of 'receptor' proteins. This opens up new questions about plant 'receptor'-ligand pairs, and PrpS-PrsS have been examined in the light of what is known about other receptors and their protein-ligand pairs in plants.

Identification of the pollen self-incompatibility determinant in Papaver rhoeas.

Higher plants produce seed through pollination, using specific interactions between pollen and pistil. Self-incompatibility is an important mechanism used in many species to prevent inbreeding; it is controlled by a multi-allelic S locus. 'Self' (incompatible) pollen is discriminated from 'non-self' (compatible) pollen by interaction of pollen and pistil S locus components, and is subsequently inhibited. In Papaver rhoeas, the pistil S locus product is a small protein that interacts with incompatible pollen, triggering a Ca(2+)-dependent signalling network, resulting in pollen inhibition and programmed cell death. Here we have cloned three alleles of a highly polymorphic pollen-expressed gene, PrpS (Papaver rhoeas pollen S), from Papaver and provide evidence that this encodes the pollen S locus determinant. PrpS is a single-copy gene linked to the pistil S gene (currently called S, but referred to hereafter as PrsS for Papaver rhoeas stigma S determinant). Sequence analysis indicates that PrsS and PrpS are equally ancient and probably co-evolved. PrpS encodes a novel approximately 20-kDa protein. Consistent with predictions that it is a transmembrane protein, PrpS is associated with the plasma membrane. We show that a predicted extracellular loop segment of PrpS interacts with PrsS and, using PrpS antisense oligonucleotides, we demonstrate that PrpS is involved in S-specific inhibition of incompatible pollen. Identification of PrpS represents a major advance in our understanding of the Papaver self-incompatibility system. As a novel cell-cell recognition determinant it contributes to the available information concerning the origins and evolution of cell-cell recognition systems involved in discrimination between self and non-self, which also include histocompatibility systems in primitive chordates and vertebrates.

Microtubules are a target for self-incompatibility signaling in Papaver pollen.

Perception and integration of signals into responses is of crucial importance to cells. Both the actin and microtubule cytoskeleton are known to play a role in mediating diverse stimulus responses. Self-incompatibility (SI) is an important mechanism to prevent self-fertilization. SI in Papaver rhoeas triggers a Ca(2+)-dependent signaling network to trigger programmed cell death (PCD), providing a neat way to inhibit and destroy incompatible pollen. We previously established that SI stimulates F-actin depolymerization and that altering actin dynamics can push pollen tubes into PCD. Very little is known about the role of microtubules in pollen tubes. Here, we investigated whether the pollen tube microtubule cytoskeleton is a target for the SI signals. We show that SI triggers very rapid apparent depolymerization of cortical microtubules, which, unlike actin, does not reorganize later. Actin depolymerization can trigger microtubule depolymerization but not vice versa. Moreover, although disruption of microtubule dynamics alone does not trigger PCD, alleviation of SI-induced PCD by taxol implicates a role for microtubule depolymerization in mediating PCD. Together, our data provide good evidence that SI signals target the microtubule cytoskeleton and suggest that signal integration between microfilaments and microtubules is required for triggering of PCD.

Initiation of programmed cell death in self-incompatibility: role for cytoskeleton modifications and several caspase-like activities.

Programmed cell death (PCD) is an important and universal process regulating precise death of unwanted cells in eukaryotes. In plants, the existence of PCD has been firmly established for about a decade, and many components shown to be involved in apoptosis/PCD in mammalian systems are found in plant cells undergoing PCD. Here, we review work from our lab demonstrating the involvement of PCD in the self-incompatibility response in Papaver rhoeas pollen. This utilization of PCD as a consequence of a specific pollen-pistil interaction provides a very neat way to destroy unwanted 'self', but not 'non-self' pollen. We discuss recent data providing evidence for SI-induced activation of several caspase-like activities and suggest that an acidification of the cytosol may be a key turning point in the activation of caspase-like proteases executing PCD. We also review data showing the involvement of the actin and microtubule cytoskeletons as well as that of a MAPK in signalling to caspase-mediated PCD. Potential links between these various components in signalling to PCD are discussed. Together, this begins to build a picture of PCD in a single cell system, triggered by a receptor-ligand interaction.