Cytoskeletal 10 nm filaments in cells of the algal phyla Chlorophyta, Charophyta, and Chrysophyta and their developmentally regulated and species-specific association with prosomes.

10 nm diameter filaments were observed in whole-mount preparations of algae of diverse phyla: Acetabularia acetabulum and A. major (Chlorophyta), Chara australis and Nitella flexilis (Charophyta), and Poterioochromonas malhamensis (Chrysophyta). A polyclonal antibody raised against a basic, 50 kDa DNA-binding protein of A. acetabulum stains the filaments of A. acetabulumand and A. major as well as of C. australis and N. flexilis. While in the perinuclear region of A. acetabulumand and A. major and throughout the cytoplasm of P. malhamensis the 10 nm filaments have a smooth appearance, in the stalk of A. acetabulumand and A. major they are densely covered by globular structures; in C. australis and N. flexilis they are less frequently associated with such material. The morphology of a part of the globular particles is quite reminiscent of prosomes. A monoclonal antibody elicited against prosomes isolated from A. acetabulum indeed decorates the globular particles on the A. acetabulum and A. major filaments. The possible role of these filament-particle associations is discussed.

Occurrence of proteinaceous 10-nm filaments throughout the cytoplasm of algae of the order Dasycladales.

Previously, whole-mount electron microscopy of nuclei extruded together with residual cytoplasm from the rhizoids of several algal species of the order Dasycladales has revealed the occurrence of an intra- and perinuclear network of 10-nm filaments morphologically indistinguishable from that of mammalian vimentin intermediate filaments. The present investigation demonstrates the existence of a filament system throughout the cytoplasm of the rhizoid, stalk, and apical tip of these giant cells. However, while the perinuclear 10-nm filaments interconnecting the nuclear surface with a perinuclear layer of large, electron-dense bodies filled with nucleoprotein material are of smooth appearance, those continuing within and beyond the perinuclear bodies are densely covered with differently sized, globular structures and, therefore, are of a very rough appearance. The filaments in the very apical tip of the cells are mainly of the smooth type. The transition from smooth to rough filaments seems to occur in the numerous perinuclear dense bodies surrounding the large nucleus. Digestion of the rough filaments with proteinase K removes the globules from the filament surface, revealing that throughout the nonvacuolar, intracellular space the filaments have the same basic 10-nm structure. On the other hand, gold-conjugated RNase A strongly binds to the filament-attached globules but not to the smooth, perinuclear, and the proteinase K-treated, rough filaments. In addition, an antibody raised against Xp54, a highly conserved protein which in Xenopus oocytes is an integral component of stored mRNP particles, decorates the rough but not the smooth 10-nm filaments. These results support the notion that the 10-nm filament system of Dasycladales cells plays a role in the transient storage of ribonucleoprotein particles in the cytoplasm and possibly fulfils a supportive function in the actomyosin-based transport of such material to various cytological destinations.

Molecular analysis of cellulose biosynthesis in Arabidopsis.

Cellulose, an abundant, crystalline polysaccharide, is central to plant morphogenesis and to many industries. Chemical and ultrastructural analyses together with map-based cloning indicate that the RSW1 locus of Arabidopsis encodes the catalytic subunit of cellulose synthase. The cloned gene complements the rsw1 mutant whose temperature-sensitive allele is changed in one amino acid. The mutant allele causes a specific reduction in cellulose synthesis, accumulation of noncrystalline beta-1,4-glucan, disassembly of cellulose synthase, and widespread morphological abnormalities. Microfibril crystallization may require proper assembly of the RSW1 gene product into synthase complexes whereas glucan biosynthesis per se does not.