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      The perichromatin region of the plant cell nucleus is the area with the strongest co-localisation of snRNA and SR proteins

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          Abstract

          The spatial organisation of the splicing system in plant cells containing either reticular ( Allium cepa) or chromocentric ( Lupinus luteus) nuclei was studied by immunolabelling of SR proteins, snRNA, and the PANA antigen, known markers for interchromatin granule clusters in mammalian cells. Electron microscope results allowed us to determine the distribution of these molecules within the structural domains of the nucleus. Similar to animal cells, in both plant species SR proteins were localised in interchromatin granules, but contrary to animal cells contained very small amounts of snRNA. The area with the strongest snRNA and SR protein co-localisation was the perichromatin region, which may be the location of pre-mRNA splicing in the plant cell nuclei. The only observable differences in the organisation of reticular and chromocentric nuclei were the size of the speckles and the number of snRNA pools in the condensed chromatin. We conclude that, despite remarkable changes in the nuclear architecture, the organisation of the splicing system is remarkably similar in both types of plant cell nuclei.

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          The online version of this article (doi:10.1007/s00425-012-1640-z) contains supplementary material, which is available to authorized users.

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          Most cited references37

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          Cajal body-specific small nuclear RNAs: a novel class of 2'-O-methylation and pseudouridylation guide RNAs.

          Cajal (coiled) bodies are conserved subnuclear organelles that are present in the nucleoplasm of both animal and plant cells. Although Cajal bodies were first described nearly 100 years ago, their function has remained largely speculative. Here, we describe a novel class of human small nuclear RNAs that localize specifically to Cajal bodies. The small Cajal body-specific RNAs (scaRNAs) are predicted or have already been demonstrated to function as guide RNAs in site-specific synthesis of 2'-O-ribose-methylated nucleotides and pseudouridines in the RNA polymerase II-transcribed U1, U2, U4 and U5 spliceosomal small nuclear RNAs (snRNAs). Our results provide strong support for the idea that the Cajal body, this mysterious nuclear organelle, provides the cellular locale for post-transcriptional modification of spliceosomal snRNAs.
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            The dynamics of a pre-mRNA splicing factor in living cells.

            Pre-mRNA splicing is a predominantly co-transcriptional event which involves a large number of essential splicing factors. Within the mammalian cell nucleus, most splicing factors are concentrated in 20-40 distinct domains called speckles. The function of speckles and the organization of cellular transcription and pre-mRNA splicing in vivo are not well understood. We have investigated the dynamic properties of splicing factors in nuclei of living cells. Here we show that speckles are highly dynamic structures that respond specifically to activation of nearby genes. These dynamic events are dependent on RNA polymerase II transcription, and are sensitive to inhibitors of protein kinases and Ser/Thr phosphatases. When single genes are transcriptionally activated in living cells, splicing factors leave speckles in peripheral extensions and accumulate at the new sites of transcription. We conclude that one function of speckles is to supply splicing factors to active genes. Our observations demonstrate that the interphase nucleus is far more dynamic in nature than previously assumed.
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              Analysis of the RNA-recognition motif and RS and RGG domains: conservation in metazoan pre-mRNA splicing factors.

              We present a systematic analysis of sequence motifs found in metazoan protein factors involved in constitutive pre-mRNA splicing and in alternative splicing regulation. Using profile analysis we constructed a database enriched in protein sequences containing one or more presumptive copies of the RNA-recognition motif (RRM). We provide an accurate alignment of RRMs and structure-based criteria for identifying new RRMs, including many that lack the prototype RNP-1 submotif. We present a comprehensive table of 125 sequences containing 252 RRMs, including 22 previously unreported RRMs in 17 proteins. The presence of a putative RRM in these proteins, which are implicated in a variety of cellular processes, strongly suggests that their function involves binding to RNA. Unreported homologies in the RRM-enriched database to the metazoan SR family of splicing factors are described for an Arg-rich human nuclear protein and two yeast proteins (S. pombe mei2 and S. cerevisiae Npl3). We have rigorously tested the phylogenetic relationships of a large sample of RRMs. This analysis indicates that the RRM is an ancient conserved region (ACR) that has diversified by duplication of genes and intragenic domains. Statistical analyses and classification of repeated Arg-Ser (RS) and RGG domains in various protein splicing factors are presented.
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                Author and article information

                Contributors
                +48-56-6114454 , +48-56-6114772 , janiaszn@umk.pl
                Journal
                Planta
                Planta
                Planta
                Springer-Verlag (Berlin/Heidelberg )
                0032-0935
                1432-2048
                24 April 2012
                24 April 2012
                August 2012
                : 236
                : 2
                : 715-726
                Affiliations
                [1 ]Department of Cell Biology, Institute of General and Molecular Biology, Nicolaus Copernicus University, Ul. Gagarina 9, 87-100 Torun, Poland
                [2 ]Department of Plant Physiology and Biotechnology, Nicolaus Copernicus University, 87-100 Torun, Poland
                [3 ]Department of Pathology, USC Keck School of Medicine, Los Angeles, CA USA
                [4 ]Division of Inflammation Biology, La Jolla Institute for Allergy and Immunology, La Jolla, CA 92037 USA
                Article
                1640
                10.1007/s00425-012-1640-z
                3404291
                22526497
                7d9b2e93-5a48-485f-97bc-8d5ce61ff9c1
                © The Author(s) 2012
                History
                : 25 October 2011
                : 21 March 2012
                Categories
                Original Article
                Custom metadata
                © Springer-Verlag 2012

                Plant science & Botany
                snrna,sr proteins,pana antigen,architecture of plant cell nucleus,splicing
                Plant science & Botany
                snrna, sr proteins, pana antigen, architecture of plant cell nucleus, splicing

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