Most of the nuclear genome of warm-blooded vertebrates is a mosaic of very long (much greater than 200 kilobases) DNA segments, the isochores; these isochores are fairly homogeneous in base composition and belong to a small number of major classes distinguished by differences in guanine-cytosine (GC) content. The families of DNA molecules derived from such classes can be separated and used to study the genome distribution of any sequence which can be probed. This approach has revealed (i) that the distribution of genes, integrated viral sequences, and interspersed repeats is highly nonuniform in the genome, and (ii) that the base composition and ratio of CpG to GpC in both coding and noncoding sequences, as well as codon usage, mainly depend on the GC content of the isochores harboring the sequences. The compositional compartmentalization of the genome of warm-blooded vertebrates is discussed with respect to its evolutionary origin, its causes, and its effects on chromosome structure and function.

CBF͞DREB1 (C-repeat-binding factor͞dehydration responsive element-binding factor 1) genes encode a small family of transcriptional activators that have been described as playing an important role in freezing tolerance and cold acclimation in Arabidopsis. To specify this role, we used a reverse genetic approach and identified a mutant, cbf2, in which the CBF2͞DREB1C gene was disrupted. Here, we show that cbf2 plants have higher capacity to tolerate freezing than WT ones before and after cold acclimation and are more tolerant to dehydration and salt stress. All these phenotypes correlate with a stronger and more sustained expression of CBF͞DREB1-regulated genes, which results from an increased expression of CBF1͞DREB1B and CBF3͞DREB1A in the mutant. In addition, we show that the expression of CBF1͞DREB1B and CBF3͞DREB1A in response to low temperature precedes that of CBF2͞DREB1C. These results indicate that CBF2͞DREB1C negatively regulates CBF1͞DREB1B and CBF3͞ DREB1A, ensuring that their expression is transient and tightly controlled, which, in turn, guarantees the proper induction of downstream genes and the accurate development of Arabidopsis tolerance to freezing and related stresses.F reezing temperatures greatly limit the geographical distribution and growing season of plants and cause negative effects on crop quality and productivity. As a consequence, appreciable effort has been conducted to determine the adaptive mechanisms plants have evolved to survive this adverse environmental condition. Many plants, including Arabidopsis, increase their freezing tolerance in response to low, nonfreezing temperatures. This process, called cold acclimation (1), is complex and involves a number of biochemical and physiological changes, ranging from alterations in lipid composition to accumulation of sugars (2). Different studies have suggested that low-temperature-regulated gene expression is critical in plants for cold acclimation (2). Low-temperature-responsive genes encode a diverse number of proteins, including enzymes involved in respiration and metabolism of carbohydrates, lipids, phenylpropanoids and antioxidants, molecular chaperones, antifreeze proteins, among others, with a believed function in freezing tolerance (2).During the past few years, substantial progress has been made toward understanding how low temperatures regulate gene expression. In particular, a family of transcription factors in Arabidopsis known either as C-repeat-binding factor (CBF)1, CBF2, and CBF3 (3, 4) or dehydration-responsive elementbinding factor (DREB)1B, DREB1C, and DREB1A (5), respectively, has been identified. These factors belong to the Apetala 2͞ethylene-responsive element-binding protein (AP2͞ EREBP) family of DNA-binding proteins (6) and bind to the cold-and dehydration-responsive DNA regulatory element (DRE) (7), also termed C-repeat (CRT) (8). CRT͞DRE elements contain the conserved CCGAC core sequence, which is sufficient to induce gene transcription under cold stress (7,8) and is present in the promoters of many cold-induc...

We have identified two genes from Arabidopsis that show high similarity with CBF1, a gene encoding an AP2 domain-containing transcriptional activator that binds to the low-temperatureresponsive element CCGAC and induces the expression of some cold-regulated genes, increasing plant freezing tolerance. These two genes, which we have named CBF2 and CBF3, also encode proteins containing AP2 DNA-binding motifs. Furthermore, like CBF1, CBF2 and CBF3 proteins also include putative nuclear-localization signals and potential acidic activation domains. The CBF2 and CBF3 genes are linked to CBF1, constituting a cluster on the bottom arm of chromosome IV. The high level of similarity among the three CBF genes, their tandem organization, and the fact that they have the same transcriptional orientation all suggest a common origin. CBF1, CBF2, and CBF3 show identical expression patterns, being induced very rapidly by low-temperature treatment. However, in contrast to most of the cold-induced plant genes characterized, they are not responsive to abscisic acid or dehydration. Taken together, all of these data suggest that CBF2 and CBF3 may function as transcriptional activators, controlling the level of low-temperature gene expression and promoting freezing tolerance through an abscisic acid-independent pathway.

Certain plants increase their freezing tolerance in response to low nonfreezing temperatures, an adaptive process named cold acclimation. Light has been shown to be required for full cold acclimation, although how light and cold signals integrate and cross-talk to enhance freezing tolerance still remains poorly understood. Here, we show that HY5 levels are regulated by low temperature transcriptionally, via a CBF-and ABA-independent pathway, and posttranslationally, via protein stabilization through nuclear depletion of COP1. Furthermore, we demonstrate that HY5 positively regulates cold-induced gene expression through the Z-box and other cisacting elements, ensuring the complete development of cold acclimation. These findings uncover unexpected functions for HY5, COP1, and the Z-box in Arabidopsis response to low temperature, provide insights on how cold and light signals integrate to optimize plant survival under freezing temperatures, and reveal the complexity of the molecular mechanisms plants have evolved to respond and adapt to their fluctuating natural environment.