A conserved family of WD-40 proteins binds to the retinoblastoma protein in both plants and animals.

In mammalian cells, the retinoblastoma (RB) protein regulates G1 progression and functions through its association with various cellular proteins. Two closely related mammalian RB binding proteins, RbAp48 and RbAp46, share sequence homology with the Msi1 protein of yeast. MSI1 is a multicopy suppressor of a mutation in the IRA1 gene involved in the Ras-cAMP pathway that regulates cellular growth. Human RbAp48 is present in protein complexes involved in histone acetylation and chromatin assembly. We report the cloning of cDNAs encoding four plant RbAp48- and Msi1-like proteins: one from tomato, LeMSI1, and three from Arabidopsis. Complementation studies confirm that LeMSI1 can function as a multicopy suppressor of the yeast ira1 mutant phenotype. The LeMSI1 protein localizes to the nucleus and binds to a 65-kD protein in wild-type as well as ripening inhibitor (rin) and Neverripe (Nr) tomato fruit. LeMSI1 also binds to the human RB protein and the RB-like RRB1 protein from maize, indicating that this interaction is conserved between plants and animals.

RRB1 and RRB2 encode maize retinoblastoma-related proteins that interact with a plant D-type cyclin and geminivirus replication protein.

Unlike mammalian and yeast cells, little is known about how plants regulate G1 progression and entry into the S phase of the cell cycle. In mammalian cells, a key regulator of this process is the retinoblastoma tumor suppressor protein (RB). In contrast, G1 control in Saccharomyces cerevisiae does not utilize an RB-like protein. We report here the cloning of cDNAs from two Zea mays genes, RRB1 and RRB2, that encode RB-related proteins. Further, RRB2 transcripts are alternatively spliced to yield two proteins with different C termini. At least one RRB gene is expressed in all the tissues examined, with the highest levels seen in the shoot apex. RRB1 is a 96-kDa nuclear protein that can physically interact with two mammalian DNA tumor virus oncoproteins, simian virus 40 large-T antigen and adenovirus E1A, and with a plant D-type cyclin. These associations are abolished by mutation of a conserved cysteine residue in RRB1 that is also essential for RB function. RRB1 binding potential is also sensitive to deletions in the conserved A and B domains, although differences exist in these effects compared to those of human RB. RRB1 can also bind to the AL1 protein from tomato golden mosaic virus (TGMV), a protein which is essential for TGMV DNA replication. These results suggest that G1 regulation in plant cells is controlled by a mechanism which is much more similar to that found in mammalian cells than that in yeast.

Replacement of the phospholipid-anchor in the contact site A glycoprotein of D. discoideum by a transmembrane region does not impede cell adhesion but reduces residence time on the cell surface.

The contact site A (csA) glycoprotein of Dictyostelium discoideum, a cell adhesion molecule expressed in aggregating cells, is inserted into the plasma membrane by a ceramide-based phospholipid (PL) anchor. A carboxyterminal sequence of 25 amino acids of the primary csA translation product proved to contain the signal required for PL modification. CsA is known to be responsible for rapid, EDTA-resistant cohesion of cells in agitated suspensions. To investigate the role of the PL modification of this protein, the anchor was replaced by the transmembrane region and short cytoplasmic tail of another plasma membrane protein of D. discoideum. In cells transformed with appropriate vectors, PL-anchored or transmembrane csA was expressed under the control of an actin promoter during growth and development. The transmembrane form enabled the cells to agglutinate in the presence of shear forces, similar to the PL-anchored wild-type form. However, the transmembrane form was much more rapidly internalized and degraded. In comparison to other cell-surface glycoproteins of D. discoideum the internalization rate of the PL-anchored csA was extremely slow, most likely because of its exclusion from the clathrin-mediated pathway of pinocytosis. Thus, our results indicate that the phospholipid modification is not essential for the csA-mediated fast type of cell adhesion but guarantees long persistence of the protein on the cell surface.

Rehydration induces rapid onset of lipid biosynthesis in desiccated Nostoc commune (Cyanobacteria).

Water, which contained [1,3-3H]glycerol, [35S]sodium sulfate, or [32P]sodium orthophosphate, was used to rehydrate air-dried cells of the desiccation-tolerant filamentous cyanobacterium Nostoc commune. The cells retained their capacities for the uptake and transport of all three compounds and, in response to rewetting, they mobilized the radiolabels into lipid precursors and initiated complex lipid biosynthesis. The onset of these events, measured in short-term, long-term and pulse-chase labeling experiments, was judged to be very rapid. The radiolabeled pool sizes of the major membrane species phosphatidylglycerol (PG) and sulfoquinovosyl diacylglycerol (SQDG) reached steady-state within several minutes, while those of the two abundant membrane glycolipids, mono- and di-glycosyldiacylglycerol (MGDG, DGDG), achieved uniform labeling within 2 h. The pattern of sulfolipid synthesis was generally more complex than the other lipid species. Analysis of the maturation of SQDG through differential labeling provided the only example of a lag in lipid maturation during the early stages (minutes) of cell rehydration. In this instance, the lag appeared to be associated specifically with the incorporation of 35SO3- by the sulfoquinovose. During the initial 2 h of rewetting there was complete turnover of 3H-label in the pools of the principal lipid precursors 1,2-sn-diacylglycerol and 1,3-diacylglycerol. In contrast, the accumulation of label by the major lipid of the heterocyst cell-wall, a non-saponifiable glycolipid, became detectable only after 24 h of rewetting. The present data are discussed in relation to the basis for desiccation tolerance in N. Commune.