Addendum to the main text (i) ATE1-/-embryos from ATE1 +/-intercrosses were present at the expected (~25%) frequency up to ~E13.5, but virtually no ATE1 -/-embryos were recovered alive by E17. Specifically, no ATE1 -/-mice were recovered amongst either 954 F 2 -generation pups of the C57BL/6J-129SvEv (mixed) background or 267 F 2 -generation pups of the 129SvEv (inbred) background. Timed intercrosses of ATE1 +/-mice were used to determine that ATE1 -/-embryos were present at approximately the expected (25%) frequency up to ~E13.5, but no ATE1 -/-embryos were recovered alive by E17.Until E12.5, ATE1 -/-embryos appeared to be morphologically normal; however, their growth stopped during E13.5-E15.5. By E14.5-E15.5, ~50% of ATE1 -/-embryos were still alive, but growth-retarded. Live E14.5-E15.5 embryos were capable of opening their mouths and flexing their bodies, suggesting the absence of gross neuromuscular defects. Sections through E13.5 ATE1-/-embryos indicated the presence and apparently normal appearance of major organs, except for the phenotypes described in the main text and below.(ii) We examined the expression of ATE1 mRNA during embryogenesis using Northern hybridization with total RNA from +/+ embryos ranging in age from E4.5 to E18.5. ATE1 mRNA was present at least as early as E4.5, and a strong spike of ATE1 expression was observed during E7.5-9.5 (Fig. S1E). The ~2 kb ATE1 transcript detected in adult mouse testis (1) was also clearly present during the spike of ATE1 expression in E7.5-E9.5 embryos (Fig. S1E). The ATE1 -allele was marked with NLS-β-galactosidase (hereafter βgal), expressed from the ATE1 promoter

proteolysis ͉ N-end rule ͉ autoinhibition ͉ E3 ͉ yeast

Plants maintain the ability to form lateral appendages throughout their life cycle and form leaves as the principal lateral appendages of the stem. Leaves initiate at the peripheral zone of the shoot apical meristem and then develop into flattened structures. In most plants, the leaf functions as a solar panel, where photosynthesis converts carbon dioxide and water into carbohydrates and oxygen. To produce structures that can optimally fulfill this function, plants precisely control the initiation, shape, and polarity of leaves. Moreover, leaf development is highly flexible but follows common themes with conserved regulatory mechanisms. Leaves may have evolved from lateral branches that are converted into determinate, flattened structures. Many other plant parts, such as floral organs, are considered specialized leaves, and thus leaf development underlies their morphogenesis. Here, we review recent advances in the understanding of how three-dimensional leaf forms are established. We focus on how genes, phytohormones, and mechanical properties modulate leaf development, and discuss these factors in the context of leaf initiation, polarity establishment and maintenance, leaf flattening, and intercalary growth.