The Arabidopsis mutant early in short days4 ( esd4 ) shows extreme early flowering and alterations in shoot development. We have identified ESD4 and demonstrate that it encodes a nuclear protein located predominantly at the periphery of the nucleus. ESD4 contains a segment of Ͼ 200 amino acids with strong similarity to yeast and animal proteases that are specific for the protein modifier SMALL UBIQUITIN-RELATED MODIFIER (SUMO). ESD4 shows a similar function to these proteases in vitro and processes the precursor of Arabidopsis SUMO (AtSUMO) to generate the mature form. This activity of ESD4 is prevented by mutations that affect the predicted active site of the protease or the cleavage site of the AtSUMO precursor. In yeast, these proteases also recycle SUMO from conjugates, and this appears to be the major role of ESD4 in vivo. This is suggested because esd4 mutants contain less free AtSUMO and more SUMO conjugates than wild-type plants, and a transgene expressing mature SUMO at high levels enhanced aspects of the esd4 phenotype. ESD4 defines an important role for protein modification by AtSUMO in the regulation of flowering.

The expression of CHALCONE SYNTHASE (CHS) expression is an important control step in the biosynthesis of flavonoids, which are major photoprotectants in plants. CHS transcription is regulated by endogenous programs and in response to environmental signals. Luciferase reporter gene fusions showed that the CHS promoter is controlled by the circadian clock both in roots and in aerial organs of transgenic Arabidopsis plants. The period of rhythmic CHS expression differs from the previously described rhythm of chlorophyll a/b-binding protein (CAB) gene expression, indicating that CHS is controlled by a distinct circadian clock. The difference in period is maintained in the wild-type Arabidopsis accessions tested and in the de-etiolated 1 and timing of CAB expression 1 mutants. These clock-affecting mutations alter the rhythms of both CAB and CHS markers, indicating that a similar (if not identical) circadian clock mechanism controls these rhythms. The distinct tissue distribution of CAB and CHS expression suggests that the properties of the circadian clock differ among plant tissues. Several animal organs also exhibit heterogeneous circadian properties in culture but are believed to be synchronized in vivo. The fact that differing periods are manifest in intact plants supports our proposal that spatially separated copies of the plant circadian clock are at most weakly coupled, if not functionally independent. This autonomy has apparently permitted tissue-specific specialization of circadian timing.Light is a key environmental signal for plants, regulating gene expression and development (Neff et al., 2000). Changes in fluence rate and light quality can occur unpredictably and rapidly during the day but have an underlying day-night cycle. Plants have evolved a circadian timing system that allows the anticipation of this predictable rhythm. When plants are deprived of environmental time cues and placed in constant ("free running") environmental conditions, circadian rhythms persist with a period of around 24 h, often for many days (Millar, 1999; McClung, 2000;Murtas and Millar, 2000;Johnson, 2001). Within the circadian system of the whole organism, the term "circadian oscillator" has been used to denote the parts of the system responsible for rhythm generation. Light-dark signals entrain the oscillator via input phototransduction pathways, synchronizing its phase with the environmental light-dark cycle and also affecting its period. Rhythmic output from the oscillator controls a large number of physiological processes in plants ( Lumsden and Millar, 1998). The abundance of 2% to 6% of RNA transcripts in Arabidopsis plants was scored as circadian-regulated in two recent microarray analyses, for example (Harmer et al., 2000;Schaffer et al., 2001).The rhythmic expression of chlorophyll a/bbinding protein (CAB or Light-Harvesting Complex [LHCB]) genes has often been used as a marker for circadian regulation in plants (for review, see Fejes and Nagy, 1998), especially using firefly (Photinus pyralis) luciferase (LUC) repor...

SUMMARYearly in short days 4, a mutation in Arabidopsis that causes early flowering and reduces the mRNA abundance of the floral repressor FLC

No abstract

One of the major properties of the semi-synthetic minimal cell, as a model for early living cells, is the ability to self-reproduce itself, and the reproduction of the boundary layer or vesicle compartment is part of this process. A minimal bio-molecular mechanism based on the activity of one single enzyme, the FAS-B (Fatty Acid Synthase) Type I enzyme from Brevibacterium ammoniagenes, is encapsulated in 1-palmitoyl-2oleoyl-sn-glycero-3-phosphatidylcholine (POPC) liposomes to control lipid synthesis. Consequently molecules of palmitic acid released from the FAS catalysis, within the internal lumen, move toward the membrane compartment and become incorporated into the phospholipid bilayer. As a result the vesicle membranes change in lipid composition and liposome growth can be monitored. Here we report the first experiments showing vesicles growth by catalysis of one enzyme only that produces cell boundary from within. This is the prototype of the simplest autopoietic minimal cell.