Demonstration of a dynamic, transcription-dependent organization of pre-mRNA splicing factors in polytene nuclei.

Baurén, Göran; Jiang, Wei‐Qin; Bernholm, Kerstin; Gu, Feng; Wieslander, Lars

doi:10.1083/jcb.133.5.929

Cited by 38 publications

(37 citation statements)

References 63 publications

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“…Consistent with previous evidence for cotranscriptional processing of pre-mRNA obtained in noninfected cells (Beyer and Osheim 1988;LeMaire and Thummel 1990;Wu et al 1991;Xing et al 1993;Bauren and Wieslander 1994;Zhang et al 1994;Tennyson et al 1995;Bauren et al 1996;Huang and Spector 1996;Neugebauer and Roth 1997), in situ evidence suggests that splicing of Ad mRNAs occurs cotranscriptionally. Namely, spliceosome components including snRNPs and non-snRNP splicing proteins are recruited to the sites of viral mRNA synthesis (Pombo et al 1994;Bridge et al 1995;Gama-Carvalho et al 1997, 2003.…”

Section: Resultssupporting

confidence: 85%

In vivo recruitment of exon junction complex proteins to transcription sites in mammalian cell nuclei

Custódio

Carvalho²,

Condado³

et al. 2004

RNA

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Section: Resultssupporting

confidence: 85%

In vivo recruitment of exon junction complex proteins to transcription sites in mammalian cell nuclei

Custódio

Carvalho²,

Condado³

et al. 2004

RNA

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“…Osheim et al 1985;Kiseleva et al 1994;Bauren et al 1996) support a model in which splicing factors accumulate at endogenous sites of transcription where splicing occurs. The detection of a high concentration of any given factor at a gene is likely to depend on the affinity of the factor for the RNA synthesized at the gene.…”

Section: Discussionmentioning

confidence: 99%

“…This has led to the suggestion that speckles contain the highest concentrations of splicing factors--perhaps serving as storage or assembly sites--and that lower concentrations may be present where splicing occurs (Fakan et al 1984;Fakan 1994;Xing et al 1995;Huang and Spector 1996). Interestingly, SR proteins and splicing snRNPs are detected at active sites of transcription on polytene and lampbrush chromosomes, which are particularly well-suited for cytological observation (Roth et al 1989(Roth et al , 1991Champlin et al 1991;Amero et al 1992;Kim et al 1992;Wu et al 1993;Bauren et al 1996). Although splicing factors accumulate at very large and highly active viral transcription units in infected mammalian cells (cf.…”

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confidence: 99%

Distribution of pre-mRNA splicing factors at sites of RNA polymerase II transcription.

Neugebauer¹,

Roth²

1997

Genes Dev.

132

118

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If pre-mRNA splicing begins during RNA synthesis, then transcriptionally active genes may be expected to contain high concentrations of pre-mRNA splicing factors. However, previous studies have localized splicing factors to a network of "speckles," which is distinct from individual sites of gene transcription where pre-mRNA is spliced. Speckles have been detected with antibodies specific for splicing snRNPs and members of the SR family of splicing factors. Here we report that dilution of these probes results in the visualization of hundreds of sites throughout the HeLa cell nucleus, the size and distribution of which are consistent with transcription units viewed with light microscopy. Importantly, these sites of highest SR protein concentration frequently coincide in three-dimensional space with active sites of RNA polymerase II transcription. A newly developed reagent specific for a single member of the SR family, SRp20, detects a subset (-20%) of these sites, suggesting the gene-specific accumulation of these splicing regulators, which have distinct functions in pre-mRNA splicing. These observations question the view that the nucleus and its functions are highly compartmentalized; instead, they support a model in which the localization of these and possibly other gene regulators is determined primarily by their function.[Key Words: Pre-mRNA splicing; RNA polymerase II; SR proteins; nuclear organization] Received December 6, 1996; revised version accepted March 18, 1997.As genes are transcribed, a diverse set of proteins and ribonucleoprotein particles (RNPs) associate with the nascent RNA. The binding of particular factors is likely to determine how the transcript will be processed to mature RNA. The observation that pre-rRNA processing and pre-mRNA splicing often occur at or very near sites of RNA synthesis (Penman et al. 1966;Rosbash and Singer 1993;Mattaj 1994) leads to the prediction that the factors regulating RNA processing may be similarly concentrated. The factors involved in pre-rRNA processing have been shown to be localized to the nucleolus, the site of rDNA transcription (Carmo-Fonseca et al. 1991;Jimenez-Garcia et al. 1994;Matera et al. 1994). In contrast, factors essential for the splicing of mRNA precursors in mammalian cell nuclei have been observed in large structures, termed "speckles," which are morphologically distinct from sites of RNA polymerase II (Pol II) transcription (Fu and Maniatis 1990;Spector 1990;Wansink et al. 1993;Zhang et al. 1994). As a first step toward resolving this enigma, we have re-examined the distribution of pre-mRNA splicing factors with respect to RNA polymerase II transcription, focusing on members of the SR protein family. 3Corresponding author. E-MAIL neugebau@embl-heidelberg.de; FAX 49 6221 387 518.Pre-mRNA splicing takes place within the spliceosome, a multicomponent complex of small nuclear ribonucleoprotein particles (snRNPs) and a number of nonsnRNP protein factors (Moore et al. 1993). The nine SR proteins constitute a family of nuclear phosphoproteins esse...

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“…SnRNP and several nonsnRNP splicing factors accumulate at both perichromatin fibrils and interchromatin granules, raising the possibility that splicing of precursor RNA may occur at these locations. Previous work suggests that some splicing of transcripts occurs at the site of transcription (Beyer and Osheim, 1988;Zhang et al, 1994;Baur6n and Wieslander, 1994;Baur6n et al, 1996;Huang and Spector, 1996); but splicing has also been observed to occur posttranscriptionally (Baur6n and Wieslander, 1994). Although splicing factors are present in the coiled body, this structure is not thought to be directly involved in the splicing of RNA, but may rather be involved in some aspect of the trafficking of splicing factors .…”

mentioning

confidence: 99%

Spliced exons of adenovirus late RNAs colocalize with snRNP in a specific nuclear domain.

Bridge

Riedel

Johansson

et al. 1996

The Journal of Cell Biology

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Abstract. Posttranscriptional steps in the production of mRNA include well characterized polyadenylation and splicing reactions, but it is also necessary to understand how RNA is transported within the nucleus from the site of its transcription to the nuclear pore, where it is translocated to the cytoplasmic compartment. Determining the localization of RNA within the nucleus is an important aspect of understanding RNA production and may provide clues for investigating the trafficking of RNA within the nucleus and the mechanism for its export to the cytoplasm. We have previously shown that late phase adenovirus-infected cells contain large clusters of snRNP and non-snRNP splicing factors; the presence of these structures is correlated with high levels of viral late gene transcription. The snRNP clusters correspond to enlarged interchromatin granules present in late phase infected cells. Here we show that polyadenylated RNA and spliced tripartite leader exons from the viral major late transcription unit are present in these same late phase snRNP-containing structures. We find that the majority of the steady state viral RNA present in the nucleus is spliced at the tripartite leader exons. Tripartite leader exons are efficiently exported from the nucleus at a time when we detect their accumulation in interchromatin granule clusters. Since the enlarged interchromatin granules contain spliced and polyadenylated RNA, we suggest that viral RNA may accumulate in this late phase structure during an intranuclear step in RNA transport.T HE production of eukaryotic mRNAs can be considered in several steps; these include transcription, polyadenylation, splicing, and export of mRNA from the nucleus to the cytoplasm. Export involves movement of the RNA within the nucleus from the site of transcription to the nuclear pore, followed by transport of the message through the nuclear pore and into the cytoplasm. Although it is convenient to think of these posttranscriptional processes as separate steps, they are likely to be inter-related during the production of RNA in vivo. All of these reactions occur within the structural framework of the nucleus; thus the nuclear organization of gene expression activities has recently received considerable attention (for reviews see Carter, 1994;Wansink et al., 1994;.The nucleus has a high degree of organization with the nucleolus as the most obvious and well characterized nuclear compartment (Scheer et al., 1993 Fakan, 1994). These include perichromatin fibrils, interchromatin granules, and coiled bodies. Perichromatin fibrils are strongly labeled after short pulses of [3H]uridine indicating that transcription occurs at these structures. Perichromatin fibrils are thought to be the in situ forms of newly synthesized hnRNA. Interchromatin granules are not strongly labeled by a pulse of [3H]uridine and are therefore not likely to be locations of transcription, but several studies have shown that polyadenylated RNA is present in these structures (Carter et al., 1991(Carter et al., , 1993Visa et al....

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Demonstration of a dynamic, transcription-dependent organization of pre-mRNA splicing factors in polytene nuclei.

Cited by 38 publications

References 63 publications

In vivo recruitment of exon junction complex proteins to transcription sites in mammalian cell nuclei

In vivo recruitment of exon junction complex proteins to transcription sites in mammalian cell nuclei

Distribution of pre-mRNA splicing factors at sites of RNA polymerase II transcription.

Spliced exons of adenovirus late RNAs colocalize with snRNP in a specific nuclear domain.

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