Cytokinin metabolism and signal transduction

Chapter 4. Cytokonin metabolism and signal transduction

4.1 Introduction 4.2 Cytokonin metabolism 4.2.1 Cytokinin biosynthesis 4.2.2 Cytokinin interconversion and conjugation 4.2.3 Cytokinin catabolism 4.3 Cytokinin signal transduction 4.3.1 Cytokinin signal perception 4.3.2 Cytokinin signal transduction 4.3.2.1 Histidine phospho-transfer proteins 4.3.2.2 Response regulators 4.3.2.2.1 B-type response regulators 4.3.2.2.2 A-type response regulators 4.4 Conclusions

Cytokinins
Cytokinins (CK) are a class of plant growth substances (plant hormones) active in promoting cell division, and are also involved in cell growth, differentiation, and other physiological processes. There are two kinds of cytokinins: Adenin cytokinins Examples: kinetin, zeatin, benzyl adenine ( same as 6- Benzylaminopurine). The DNA base adenine is a structural analogue of cytokinins and have low cytokinin bioactivity. Phenylurea cytokinins Example: N, N'-diphenylurea Although their chemical compositions differ, there is a structural correlation between adenine cytokinins and urea cytokinin, and both show similar biological activities.

Cytokinin signal transduction via a His-to-Asp phosphorelay

< Two-component system(TCS) > 1.Membrane bound receptor kinase : *Senses the signal and autophosphorylates 2.Response regulator : *Phosphorylation by the receptor kinase *Activates the transcription of its target genes *Initiates another output reaction

Current Opinion in Plant Biology 2003, 6:480488

Structures of cytokinin receptors

Cytokinin receptors consist of input domain, histidine kinase, receiver domain.

AHKs can interact with all five AHPs

Fig. 1 Both the Arabidopsis His-kinase AHK4 and HPt factor AHP2 function in E. coli. When the Arabidopsis AHK4 gene is expressed in this particular mutant, AHK4 is capable of signaling the downstream YojN RcsB csp::lacZ pathway in response to external cytokinins. Consequently, the target csp::lacZ gene is activated. However, when the Arabidopsis AHP2 gene is co-expressed in this E. coli assay system, AHP2 can titrate a phosphoryl group form AHK4, thereby inhibiting the AHK4 YojN phosphorelay.

Plant and Cell Physiology, 2002, Vol. 43, No. 1 123-129

Ligand specificity of cytokinin receptors

Fig 1.

Fig. 1 Comparison of the sensitivities of CRE1/AHK4 and AHK3 to different cytokinin bases in the E. coli assay.

Plant Cell Physiol. 45(9): 12991305 (2004)

Cytokinin receptors have at least a partial overlap in their function

Figure 1. Shoot Development of ahk Mutant Plants.

The Plant Cell, Vol. 18, 4054, January 2006

Cytokinin receptors have at least a partial overlap in their function

Figure 8. Contributions of Different Cytokinin Receptors and Receptor Combinations to Cytokinin-Regulated Processes.

The Plant Cell, Vol. 18, 4054, January 2006

Arabidopsis Histidine phospho-transfer proteins(AHP)

Current Opinion in Plant Biology 2003, 6:480488

Histidine phospho-transfer proteins

The central role of the AHPs as mediators of the cytokinin signal requires that they can interact with AHKs and ARRs.

AHP2, AHP3, AHP5 : Expressed all tissues AHP1 : Roots AHP4 : Shoot tissues Bioscience, Biotechnology, and Biochemistry Vol. 68 (2004) , No. 2 pp.462-465

AHP proteins as cytoplasm and nucleus shuttles

Figure 2 AHP acts as a shuttle between the cytoplasm and nucleus in cytokinin signalling.

NATURE |VOL 413 | 27 SEPTEMBER 2001

Arabidopsis Response Regulators(ARR)

Current Opinion in Plant Biology 2003, 6:480488

Response regulators

Analysis of the Arabidopsis genome sequence reveals the existence of 22 predicted response regulator (ARR) genes. They contain a RR domain the conserved aspartate of this domain, which is required for the His-to-Asp phospho-relay. The ARRs are divided into two major classes, the A- and the B-type, on the basis of their structure. The 11 type-A ARRs consist mainly of the receiver domain with a short extension at the N- and C-terminal ends. The 11 type-B ARRs contain a C-terminal output domain in addition to the receiver domain.

Current Opinion in Plant Biology 2003, 6:480488

B-type response regulators

ARR1/ARR2, ARR10/ARR12, ARR11,ARR14, ARR18 Expressed in almost all tissues. (ARR14 : Not found in roots ARR18 : Flower specific) ARR13/ARR21 Expressed in reproductive tissues. ARR19/ARR20 Expressed in reproductive tissues.

Figure 7. Overview of Arabidopsis type-B ARR localization based on GUS analysis.

Plant Physiol. Vol. 135, 2004

Subcellular localization of B-type ARRs

Localize to nucleus. Predicted role as transcription factors.

Figure 6. Nuclear localization of type-B ARRs.

*Several studies reporting B-type ARR gene loss of function mutants did not display any phenotypical alteration, except for longer roots. Redundant B-type ARR functions *Overexpression of ARR2 did not cause dramatic phenotipic changes. *Substitution of the conserved Asp80 with Glu created a dominant-active form of ARR2, whose overexpression caused severe pleiotropic growth aberrations, including disturbance in the shoot meristem and an aberrant leaf shape.

The EMBO Journal VOL 23 | NO 16 | 2004

A-type response regulators

< Expression patterns of A-type response regulators > ARR3, ARR4 : vascular tissue of shoots and roots ARR5, ARR6 : meristems of shoots and roots ARR8, ARR9 : roots
*Several A-type ARRs have been shown to be negative regulators of cytokinin signaling in a protoplast system.(ARR3, ARR6, ARR8, ARR9) *Several A- type ARRs might be positive regulators and thus enhance the cytokinin response.

Negative feedback regulation of cytokinin signaling by A-type ARRs.

*For all A-type ARRs tested, interactions with some or all of the AHPs were detected. However no direct interaction between the A- and Btype ARRs was found. Negative regulation of the signaling pathway by A-type ARRs operates via their interaction with AHPs.

Conclusions
< Two different type of cytokinin hormonal activities exist in the plant > *A local(paracrine or autocrine) activity, which may regulate mainly cell division. *Long-distance signaling(endocrine-like), in which cytokinin may sever primarily as a root-to-shoot signal and may regulate physiological processes.

< Future goals > *To find out which are the individual functions of the cytokinin metabolism and signaling. *To determine how signal specification is generated in this partially redundant signaling system. *Integration of this signaling system into other pathways within the plant cell, thus to understand the molecular mechanisms that drive the developmental and physiological processes related to cytokinin.

Features of cytokinin receptors and components of the signalling pathway

Current Opinion in Plant Biology 2003, 6:480488