Brassinosteroids regulate plant growth and development through a protein complex that includes the leucine-rich repeat receptor-like protein kinase (LRR-RLK) brassinosteroid-insensitive 1 (BRI1). Activation tagging was used to identify a dominant genetic suppressor of bri1, bak1-1D (bri1-associated receptor kinase 1-1Dominant), which encodes an LRR-RLK, distinct from BRI1. Overexpression of BAK1 results in elongated organ phenotypes, while a null allele of BAK1 displays a semidwarfed phenotype and has reduced sensitivity to brassinosteroids (BRs). BAK1 is a serine/threonine protein kinase, and BRI1 and BAK1 interact in vitro and in vivo. Expression of a dominant-negative mutant allele of BAK1 causes a severe dwarf phenotype, resembling the phenotype of null bri1 alleles. These results indicate BAK1 is a component of BR signaling.

Stomata are specialized epidermal structures that regulate gas (CO 2 and O 2 ) and water vapor exchange between plants and their environment. In Arabidopsis thaliana, stomatal development is preceded by asymmetric cell divisions, and stomatal distribution follows the one-cell spacing rule, reflecting the coordination of cell fate specification. Stomatal development and patterning are regulated by both genetic and environmental signals. Here, we report that Arabidopsis MITOGEN-ACTIVATED PROTEIN KINASE3 (MPK3) and MPK6, two environmentally responsive mitogen-activated protein kinases (MAPKs), and their upstream MAPK kinases, MKK4 and MKK5, are key regulators of stomatal development and patterning. Loss of function of MKK4/MKK5 or MPK3/MPK6 disrupts the coordinated cell fate specification of stomata versus pavement cells, resulting in the formation of clustered stomata. Conversely, activation of MKK4/MKK5-MPK3/MPK6 causes the suppression of asymmetric cell divisions and stomatal cell fate specification, resulting in a lack of stomatal differentiation. We further establish that the MKK4/MKK5-MPK3/MPK6 module is downstream of YODA, a MAPKKK. The establishment of a complete MAPK signaling cascade as a key regulator of stomatal development and patterning advances our understanding of the regulatory mechanisms of intercellular signaling events that coordinate cell fate specification during stomatal development.

No abstract

Abscission is a developmental program that results in the active shedding of infected or nonfunctional organs from a plant body. Here, we establish a signaling pathway that controls abscission in Arabidopsis thaliana from ligand, to receptors, to downstream effectors. Loss of function mutations in Inflorescence Deficient in Abscission (IDA), which encodes a predicted secreted small protein, the receptor-like protein kinases HAESA (HAE) and HAESA-like 2 (HSL2), the Mitogen-Activated Protein Kinase Kinase 4 (MKK4) and MKK5, and a dominant-negative form of Mitogen-Activated Protein Kinase 6 (MPK6) in a mpk3 mutant background all have abscission-defective phenotypes. Conversely, expression of constitutively active MKKs rescues the abscission-defective phenotype of hae hsl2 and ida plants. Additionally, in hae hsl2 and ida plants, MAP kinase activity is reduced in the receptacle, the part of the stem that holds the floral organs. Plants overexpressing IDA in a hae hsl2 background have abscission defects, indicating HAE and HSL2 are epistatic to IDA. Taken together, these results suggest that the sequential action of IDA, HAE and HSL2, and a MAP kinase cascade regulates the programmed separation of cells in the abscission zone.protein phosphorylation ͉ signal transduction A bscission is a physiological process that involves the programmed separation of entire organs, such as leaves, petals, flowers, and fruit. Abscission allows plants to discard nonfunctional or infected organs, and promotes dispersion of progeny. At the cellular level, abscission is the hydrolysis of the middle lamella of an anatomically specialized cell layer, the abscission zone (AZ), by cell wall-modifying and hydrolyzing enzymes. Thus, abscission requires both the formation of the AZ early in the development of a plant organ and the subsequent activation of the cell separation response (1-4).Studies using Arabidopsis thaliana have implicated the involvement of several different genes in the control of abscission including potential signal molecules, receptors and other gene products (4). HAESA (HAE), one of the first Arabidopsis receptor-like protein kinases (RLK) identified, is expressed in floral organ AZs and antisense experiments show a reduction in the level of HAE protein is correlated with the degree of defective floral organ abscission. Expression of HAE is not altered in etr1-1 (an ethylene-insensitive mutation), implying an ethyleneindependent role in abscission (5). Inflorescence Deficient in Abscission (IDA) encodes a small protein with an N-terminal signal peptide. Analysis of ida mutant plants indicates IDA regulates floral organ abscission through an ethylene insensitive pathway (6). Overexpression of IDA results in early abscission and the production of a white substance in the floral AZs. The main components of the white substance are arabinose and galactose (7).Here, we report that components of a MAPK signaling cascade also have roles in the regulation of abscission. A MAPK cascade is a regulatory module with three protein kina...

CorrectionsBIOCHEMISTRY. For the article ''Arg-302 facilitates deprotonation of Glu-325 in the transport mechanism of the lactose permease from Escherichia coli'' by Miklós Sahin-Tóth and H. Ronald Kaback, which appeared in number 11, May 22, 2001, of Proc. Natl. Acad. Sci. USA (98, 6068 -6073; First Published May 15, 2001; 10.1073͞pnas.111139698), Fig. 1 was printed incorrectly due to a printer's error. The correct figure and its legend appear below.www.pnas.org͞cgi͞doi͞10.1073͞pnas.151252598 Fig. 1.Model for H ϩ translocation during lactose͞H ϩ symport via lac permease. For clarity, 6 of the 12 helices that compose the permease are shown. The gray area designates the low dielectric environment of the lipid bilayer. (A) In the ground-state conformation, the relevant H ϩ is shared by His-322 (helix X) and Glu-269 (helix VIII), whereas Arg-302 (helix IX) is charge-paired with Glu-325 (helix X). In this conformation, lac permease binds substrate with high affinity at the outer surface (S o ). Glu-126 (helix IV) and Arg-144 (helix V) are charge-paired and represent the major components of the substrate-binding site. Also shown is the charge-pair between Asp-240 (helix VII) and Lys-319 (helix X), which are not essential for the mechanism. (B) Substrate binding induces a conformational change that disrupts the E269͞H322 and R302͞E325 charge-pairs and leads to the transfer of the H ϩ to Glu-325, now stabilized by the low dielectric environment. At the same time, the substrate-binding site becomes exposed to the inner surface of the membrane (S i ). After substrate dissociation, Glu-325 deprotonates at the inside surface (because of the rejuxtaposition of Glu-325 with Arg-302) as the permease relaxes back to the ground-state conformation.