Interactions between abscisic acid and ethylene signaling cascades.

We screened for mutations that either enhanced or suppressed the abscisic acid (ABA)-resistant seed germination phenotype of the Arabidopsis abi1-1 mutant. Alleles of the constitutive ethylene response mutant ctr1 and ethylene-insensitive mutant ein2 were recovered as enhancer and suppressor mutations, respectively. Using these and other ethylene response mutants, we showed that the ethylene signaling cascade defined by the ETR1, CTR1, and EIN2 genes inhibits ABA signaling in seeds. Furthermore, epistasis analysis between ethylene- and ABA-insensitive mutations indicated that endogenous ethylene promotes seed germination by decreasing sensitivity to endogenous ABA. In marked contrast to the situation in seeds, ein2 and etr1-1 roots were resistant to both ABA and ethylene. Our data indicate that ABA inhibition of root growth requires a functional ethylene signaling cascade, although this inhibition is apparently not mediated by an increase in ethylene biosynthesis. These results are discussed in the context of the other hormonal regulations controlling seed germination and root growth.

The role of ABI1 in abscisic acid signal transduction: from gene to cell.

The semi-dominant abi1-1 mutation of Arabidopsis interferes with multiple aspects of abscisic acid signal transduction resulting in reduced seed dormancy and sensitivity of root growth in ABA. Furthermore, the mutant transpires excessively as a result of abnormal stomatal regulation leading to a wilty phenotype. The ABI1 gene has been cloned. The carboxyl-terminal domain of the predicted ABI1 protein is related to the 2C class of serine-threonine phosphatases while no overt homology was found in the extended amino terminus. A combination of in vitro assays and yeast mutant complementation studies confirmed that ABI1 is a functional protein phosphatase 2C. The abi1-1 mutation converts the amino acid glycine180 to aspartic acid, and in the above test systems, causes a partial loss of the phosphatase activity. In transgenic Nicotiana benthamiana guard cells, the abi1-1 gene causes a reduction in the background current of the outward-rectifying potassium channels, and also in the abscisic acid-sensitivity of both the outward- and the inward-rectifying potassium channels in the plasma membrane. However, normal sensitivity of both potassium channels to, and stomatal closure in, abscisic acid was recovered in the presence of H7 and staurosporine, both broad-range protein kinase antagonists. These results suggest the aberrant potassium channel behavior as a major consequence of abi1-1 action and implicate ABI1 as part of a phosphatase/kinase pathway that modulates the sensitivity of guard-cell potassium channels to abscisic acid-evoked signal cascades.

Characterization of a negative regulator of exopolysaccharide production by the plant-pathogenic bacterium Pseudomonas solanacearum.

Wild-type strains of the bacterial wilt pathogen Pseudomonas solanacearum exhibit reduced exopolysaccharide production and virulence when transformed with plasmids carrying the epsR locus. To understand the function of epsR, we used mutagenesis and DNA sequencing to identify the gene responsible for the shutoff of exopolysaccharide production. The epsR gene encodes a 236-amino-acid polypeptide that, based on polypeptide sequence homology, has significant similarity to other proteins of the luxR family of environmentally responsive, two-component regulatory systems. When a mutated copy of the epsR gene was marker-exchanged into the wild-type P. solanacearum chromosome, however, we observed no effect on growth in culture or on exopolysaccharide production. This suggests that the EpsR phenotype becomes apparent only via overproduction of the EpsR protein. By means of an antiserum directed against the EpsR protein, we detected the overproduction of EpsR in cell lysates of a strain of P. solanacearum harboring a multicopy plasmid with an active epsR gene but not in one harboring the same plasmid with a mutated epsR gene.

Human centrosomal epitope is shared specifically with human lactate dehydrogenase-B isozyme.

A rabbit serum (0013) used to identify pericentriolar proteins from isolated centrosomes (Gosti-Testu, F., Marty, M.C., Berges, J., Maunoury, R. and Bornens, M. (1986) EMBO J. 5, 2545-2550) was shown also to react through the same epitope with several non-centrosomal proteins including a major 36 kDa cytosolic antigen. This protein was identified to be human lactate dehydrogenase and the co-distribution of 0013 epitope on the centrosomal protein and on lactate dehydrogenase (LDH) was shown to be specific for human cells (Gosti, F., Marty, M.C., Courvalin, J.C., Maunoury, R. and Bornens, M. (1987) Proc. Natl. Acad. Sci. USA 84, 1000-1004). Human hepatic cells constitute, so far, the only exception to this co-distribution rule. By using this cell type which expresses only the LDH-A4 isozyme, we demonstrate that 0013 epitope is specific for the human LDH-B subunit, making serum 0013 the strongest anti-LDH-B available so far. The evolutionary and physiological significance of this situation is discussed.

Lamins A and C are not expressed at early stages of human lymphocyte differentiation.

Lamins are major proteins of the nuclear envelope that are members of the intermediate filament protein family. In vertebrates, nuclei from differentiated tissues usually contain both lamins of the A and B subtypes, while embryonic tissues contain the B-type lamin only. We have examined the composition of the nuclear lamina in human B and T lymphocytes representative of distinct stages of lymphoid differentiation. We show here that, in both cell lineages, while lamin B is constitutively expressed at all stages of differentiation, A-type lamin expression is restricted to later developmental stages.