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Myeloid leukemia factor 1 (MLF1) is associated with the development of leukemic diseases such as acute myeloid leukemia (AML) and myelodysplastic syndrome (MDS). However, information on the physiological function of MLF1 is limited and mostly derived from studies identifying MLF1 interaction partners like CSN3, MLF1IP, MADM, Manp and the 14-3-3 proteins. The 14-3-3-binding site surrounding S34 is one of the only known functional features of the MLF1 sequence, along with one nuclear export sequence (NES) and two nuclear localization sequences (NLS). It was recently shown that the subcellular localization of mouse MLF1 is dependent on 14-3-3 proteins. Based on these findings, we investigated whether the subcellular localization of human MLF1 was also directly 14-3-3-dependent. Live cell imaging with GFP-fused human MLF1 was used to study the effects of mutations and deletions on its subcellular localization. Surprisingly, we found that the subcellular localization of full-length human MLF1 is 14-3-3-independent, and is probably regulated by other as-yet-unknown proteins.
Conserved 14-3-3 proteins have been shown to play regulatory roles in eukaryotic cells, including cell cycle control and differentiation. We were interested in the possible involvement of 14-3-3 proteins in the embryogenic process of barley (Hordeum vulgare L.). Barley microspore-derived embryo development was used as a model system. Immunolocalization of three barley 14-3-3 isoforms, 14-3-3A, 14-3-3B and 14-3-3C, was carried out using isoform-specific antibodies. In immature microspore-derived embryos, 14-3-3C was specifically expressed underneath the L1 layer of the shoot apical meristem and in the scutellum. Comparative studies showed that 14-3-3C was also expressed underneath the L1 layer of the shoot apical meristem and in the scutellum of immature zygotic embryos. We further demonstrated that 14-3-3C expression is restricted to L2 layer-derived cells of in vitro shoot meristematic cultures. Our results reveal that 14-3-3C isoform tissue-specific expression is closely related to defined events during differentiation processes in embryogenesis and in vitro meristematic cultures.
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