Functional analysis of plant Mei2-like proteins : a thesis presented in partial fulfilment of the requirements for the degree of Master of Science in Biochemistry at Massey University, Palmerston North, New Zealand
Molecular techniques were used to analyse the function of a novel class of RNA-bindmg proteins in plants, termed Mei2-like. The biochemical function of this class of proteins is unclear Although the conserved presence of three RNA recognition motifs (RRMs) in all members of the family suggests the importance of an RNA binding activity, the precise biochemical mechanism by which these proteins act is unknown. Genetic and molecular analyses of the founding member of the family. Schizosaccharomyces pombe Mei2p, provide of a conceptual framework for the studies of the plant Mei2-like proteins presented here. Therefore, the aims of this thesis were to 1) study the cellular localisation of Mei2p in plant cells, and 2) deduce the functions of plant Mei2-like genes by identifying the protein(s) that physically interact(s) with Mei2-like proteins. Transient expression of GFP-fused Mei2p in onion epidermal cells was performed to show that Mei2p localised into the nucleus in the presence of meiRNA, a non-coding mRNA. Thus plants seem to share the capacity with S. pombe for meiRNA-dependent nuclear localisation of Mei2p. Moreover, intracellular localisation of one of the plant Mei2-like proteins, TERMINAL EAR-Like 2 (TEL2), was studied in onion epidermal cells. The GFP-fused TEL2 localised into the nucleus without co-expression of any special RNA, suggesting that either some RNA species that assist nuclear localisation of TEL2 are already present in onion epidermal cells, or the mechanism of intracellular localisation of TEL2 is different from Mei2p. The yeast two-hybrid system was utilised to identify protein interactors with TEL2. Six proteins were identified, including the well-studied KORRIGAN (KOR) protein. Based on the proteins identified, speculation is offered on how these proteins interact with TEL2. Since TEL genes are expressed in the central zone (CZ) of meristems, and mitotic activity of cells in the CZ is low, TEL2 may be involved in controlling cell division in the CZ via interactions with these proteins.