Understanding molecular mechanisms that control cell fate in the shoot apical meristem is a fundamental question in plant development. Genetic and molecular studies demonstrate that maize KNOTTED1 (KN1) of the TALE (3-aa acid loop extension) class of homeodomain (HD) proteins is involved in shoot apical meristem function. We show that KN1 interacts with knotted interacting protein (KIP), a BEL1-like TALE HD protein. Interaction between KN1 and KIP is mediated by conserved domains in the N termini of both proteins. The KN1 DNA-binding sequence, TGACAG(G͞C)T, was biochemically identified, and in vitro DNA-binding assays show that individually KN1 and the HD of KIP bind specifically to this motif with low affinity. The KN1-KIP complex, however, binds specifically to this DNA-binding motif with high affinity, indicating that the association of KN1 and KIP may function in transcriptional regulation.
In plants, development of the aerial shoot is controlled by a group of cells in the shoot apex called the shoot apical meristem (SAM). The SAM is the site where organogenesis is initiated and from which leaves, flowers, or other determinate structures are produced. Because plant development is a continuous process, the SAM must balance the process of differentiation and maintenance to allow growth.Maize knotted1 (kn1) and the Arabidopsis ortholog SHOOT-MERISTEMLESS (STM) are members of the knox family of genes that are required for SAM function (1). The expression of kn1 and STM is restricted to the indeterminate cells of the SAM. Moreover, this pattern of expression is excluded from the zone of the SAM that gives rise to organs (2, 3). Loss-of-function mutations in kn1 and STM produce a shootless phenotype, where the SAM prematurely terminates after initiating cotyledons, demonstrating that kn1 and STM function in meristem maintenance (3-5). In backgrounds where some shoot formation occurs, organ production is reduced, indicating that vegetative and inflorescence meristems are compromised (4,(6)(7)(8). In addition to maintenance functions, class 1 knox genes, including kn1, are capable of respecifying cell fates when ectopically expressed in the leaf (1). Studies that support this hypothesis come from analysis of dominant mutations in the maize knox genes, kn1, rough sheath1 (rs1), liguleless3 (lg3), and gnarley1 (gn1) (9-12). Each of these phenotypes is caused by ectopic leaf expression, resulting in proximal leaf cell identities transposed distally into the blade (1, 13).The knox gene family was the first group of plant genes identified that encodes homeodomain (HD) proteins (14). In animals, HD-containing proteins function as transcription factors that are important regulators of animal cell fate and development (15). The HD is a conserved structure that contains three ␣ helices, and a sequence motif in the third helix recognizes and binds to the appropriate DNA sequence (15). Amino acid sequence comparison studies have subdivided the HD family into two classes of proteins: the typical HD class and the TALE (3-aa loop...