caa a22 4500 467894744 CHVBK 20180406152759.0 cr unu---uuuuu 170328e20061101xx s 000 0 eng 10.1007/s11103-006-9046-6 doi (NATIONALLICENCE)springer-10.1007/s11103-006-9046-6 Systematic identification of factors involved in post-transcriptional processes in wheat grain [Elektronische Daten] [Sergiy Lopato, Ljudmilla Borisjuk, Andrew Milligan, Neil Shirley, Natalia Bazanova, Peter Langridge] Post-transcriptional processing of primary transcripts can significantly affect both the quantity and the structure of mature mRNAs and the corresponding protein products. It is an important mechanism of gene regulation in animals, yeast and plants. Here we have investigated the interactive networks of pre-mRNA processing factors in the developing grain of wheat (Triticum aestivum), one of the world's major food staples. As a first step we isolated a homologue of the plant specific AtRSZ33 splicing factor, which has been shown to be involved in the early stages of embryo development in Arabidopsis. Real-time PCR showed that the wheat gene, designated TaRSZ38, is expressed mainly in young, developing organs (flowers, root, stem), and expression peaks in immature grain. In situ hybridization and immunodetection revealed preferential abundance of TaRSZ38 in mitotically active tissues of the major storage organ of the grain, the endosperm. The protein encoded by TaRSZ38 was subsequently used as a starting bait in a two-hybrid screen to identify additional factors in grain that are involved in pre-mRNA processing. Most of the identified proteins showed high homology to known splicing factors and splicing related proteins, supporting a role for TaRSZ38 in spliceosome formation and 5′ site selection. Several clones were selected as baits in further yeast two-hybrid screens. In total, cDNAs for 16 proteins were isolated. Among these proteins, TaRSZ22, TaSRp30, TaU1-70K, and the large and small subunits of TaU2AF, are wheat homologues of known plant splicing factors. Several, additional proteins are novel for plants and show homology to known pre-mRNA splicing, splicing related and mRNA export factors from yeast and mammals. Springer Science+Business Media B.V., 2006 Grain development nationallicence Pre-mRNA processing factors nationallicence Protein-protein interaction nationallicence Splicing nationallicence Yeast two-hybrid system nationallicence AD : Activation domain nationallicence BD : Binding domain nationallicence DAP : Days after pollination nationallicence dbEST : GenBank expressed sequence tag database nationallicence GUS : β-Glucuronidase nationallicence NEB : Nuclei extraction buffer nationallicence NLS : Nuclear localization signal nationallicence NMD : Nonsense mediated decay nationallicence RRM : RNA recognition motif nationallicence RSZproteins : Arginine/serine proteins with one or two zinc fingers nationallicence SCL proteins : SC35-like proteins nationallicence SD domains : Serine/aspartic acid rich domains nationallicence snRNPs : Small nuclear ribonucleoprotein particles nationallicence SR domains : Serine/arginine rich domains nationallicence TRN-SR : Transportin-SR nationallicence Lopato Sergiy Australian Centre for Plant Functional Genomics, The University of Adelaide, PMB1, 5064, Glen Osmond, SA, Australia aut Borisjuk Ljudmilla Institut fur Pflanzengenetik und Kulturpflanzenforschung, D-06466, Gatersleben, Germany aut Milligan Andrew Australian Centre for Plant Functional Genomics, The University of Adelaide, PMB1, 5064, Glen Osmond, SA, Australia aut Shirley Neil Australian Centre for Plant Functional Genomics, The University of Adelaide, PMB1, 5064, Glen Osmond, SA, Australia aut Bazanova Natalia Australian Centre for Plant Functional Genomics, The University of Adelaide, PMB1, 5064, Glen Osmond, SA, Australia aut Langridge Peter Australian Centre for Plant Functional Genomics, The University of Adelaide, PMB1, 5064, Glen Osmond, SA, Australia aut Plant Molecular Biology Kluwer Academic Publishers 62/4-5(2006-11-01), 637-653 0167-4412 62:4-5<637 2006 62 11103 https://doi.org/10.1007/s11103-006-9046-6 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s11103-006-9046-6 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Lopato Sergiy Australian Centre for Plant Functional Genomics, The University of Adelaide, PMB1, 5064, Glen Osmond, SA, Australia aut NATIONALLICENCE 700 1- Borisjuk Ljudmilla Institut fur Pflanzengenetik und Kulturpflanzenforschung, D-06466, Gatersleben, Germany aut NATIONALLICENCE 700 1- Milligan Andrew Australian Centre for Plant Functional Genomics, The University of Adelaide, PMB1, 5064, Glen Osmond, SA, Australia aut NATIONALLICENCE 700 1- Shirley Neil Australian Centre for Plant Functional Genomics, The University of Adelaide, PMB1, 5064, Glen Osmond, SA, Australia aut NATIONALLICENCE 700 1- Bazanova Natalia Australian Centre for Plant Functional Genomics, The University of Adelaide, PMB1, 5064, Glen Osmond, SA, Australia aut NATIONALLICENCE 700 1- Langridge Peter Australian Centre for Plant Functional Genomics, The University of Adelaide, PMB1, 5064, Glen Osmond, SA, Australia aut NATIONALLICENCE 773 0- Plant Molecular Biology Kluwer Academic Publishers 62/4-5(2006-11-01), 637-653 0167-4412 62:4-5<637 2006 62 11103 Metadata rights reserved Springer special CC-BY-NC licence nationallicence SWISSBIB 462662101 BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer