The interactions they are a changin’ – The complex role of protein interactions in the evolution of flower development

by Aalt-Jan Van Dijk

Among the various transcription factors involved in regulating flowering, no doubt MADS domain proteins involve some of the best studied. B-class proteins are a specific type of MADS domain proteins, named after their role in the classic ABC model for floral development. According to the ABC model, B-class proteins are involved in petal and stamen formation; for more background information on this model, you might want to read the recent post by Theissen and Parcy  (which focusses on other components of the ABC model, namely A-class proteins). B-class proteins can either form heterodimers, involving two different B-class proteins, or homodimers. Variation in B-class proteins has been suggested to be relevant for variation in floral organ development. However, a lot is still unclear about how variation in these proteins and their interactions influences phenotypic differences related to flowering. Better understanding of the evolution of B-class dimerization is clearly needed. A recent paper in Molecular Biology and Evolution  (Bartlett et al., 2016) uncovers various layers of complexity of this evolution.

First, Bartlett et al., 2016 characterized obligate heterodimerization versus homodimerization in taxa spanning the Poales (the order that contains the grass family), and found multiple transitions between factultative homodimerization and obligate heterodimerization. Such evolutionary changes were also present specifically within the grasses.

The next layer of complexity is, that there is a clear context-dependence of the effect of specific amino acids on the dimerization landscape. This is demonstrated by results from the experiments presented by Bartlett et al. to find sequence regions influencing homodimerization versus heterodimerization. These experiments involved three B-class proteins, two of which formed homodimers: J-PI (from the grass relative Joinvillea) and BdSTS1 (from Brachypodium distachyon) and a third one forming heterodimers (STS1, a maize protein). See figure for a schematic overview of the main findings of these experiments.

Figure1

Context-dependence of the effect of specific domains on dimerization.
Ovals represent B-class proteins: green, B. distachyon BdSTS1; blue, maize STS1; red, Joinvillea J-PI. Dashed line indicates homodimerization. The STS1 I-domain (small blue oval) disrupts homodimerization in the context of J-PI, but not so in BdSTS1. Similarly, the J-PI I-domain (small red oval) enables homodimerization of STS1, but the BdSTS1 I-domain (small green oval) does not.

It was found that the I-domain of J-PI   is sufficient for homodimerization of STS1; here the I-domain refers to a specific domain in MADS proteins. This domain is in fact known to be relevant for MADS domain interaction specificity in general. However the I-domain of BdSTS1 was not sufficient for homodimerization of STS1. Reciprocally, the STS1 I-domain was sufficient to abolish J-PI homodimerization but did not affect BdSTS1 homodimerization.  A comparison of I-domain sequence of STS1 and J-PI showed four amino acid differences. A single amino acid change (Gly81 to Asp) was sufficient to confer homodimerization ability on STS1, and the reciprocal change of Asp to Gly prevented J-PI from homodimerization. Intriguingly, however, the homodimerizing BdSTS1 I-domain also contains Asp at position 81, but this domain was not sufficient to confer homodimerization ability to STS1. BdSTS1 contains Ile at position 73, and introducing this Ile abolished the homodimerization capacity that the Gly81Asp mutation conferred on STS1. In brief, this indicates that a specific amino acid (Asp) can be sufficient to allow homodimerization in one sequence context but not so in another sequence context. This context-dependence of the role of specific amino acids provides insight to the multiple evolutionary routes via which B-class heterodimerization versus homodimerization evolved.

It is interesting to see that two computational methods contributed to the identification of relevant amino acids (Bartlett et al., 2016). One involves the analysis of positive selection. The other involves the prediction of interaction motifs. Both approaches were used for further identification of amino acids that contribute to dimerization landscape.

A final level of complexity in their analysis is the interconnectedness of coding and non-coding changes. Note that Theissen and Parcy on their post on the floral A-function gave an example of the importance of non-coding changes. Bartlett et al. found that the (homodimerizing) J-PI and the (non-homodimerizing) STS1 both showed similar rescue of an Arabidopsis pi mutant, and both could not rescue an ap3 mutant. (PI and AP3 are the two Arabidopsis B-class proteins). However, they found a distinction between J-PI and STS1 when they were expressed at a high level in developing sepals. Only J-PI expression resulted in transformation of the first whorl organs into petals. This shows that J-PI, when expressed at a high level in a novel domain, differs from STS1 in its effect on floral development. The J-PI homodimer on its own is not sufficient to confer B-class function in Arabidopsis, but as a novel protein complex in a novel domain it can effect phenotypic change. The interactions they are a changin – but the effect of changing protein interactions clearly depends on the expression pattern of the protein in question, and of its interactors.

Reference
Bartlett M, Thompson B, Brabazon H, Del Gizzi R, Zhang T and Whipple C. 2016. Evolutionary Dynamics of Floral Homeotic Transcription Factor Protein–Protein Interactions. Molecular Biology and Evolution 33(6):1486–1501. doi:10.1093/molbev/msw031

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About Flowering Highlights

Flowering Newsletter published by the Journal of Experimental Botany
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