Structural/functional properties of a mammalian multi-component structure containing all major spliceosomal small nuclear ribonucleoprotein particles

Μωραιτου, Μαρινα; Patrinou-Georgoula, Meropi; Guialis, Apostolia

doi:10.1042/bj3320135

Cited by 4 publications

(3 citation statements)

References 37 publications

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“…As judged from the sustained presence of pre-mRNA and intermediate products, splicing had not reached completion after 60 min of incubation. As shown previously[17,28], an almost complete disappearance of intermediate products is not seen until 120 min after the start of the splicing reaction.InFigure 1(B), the RNA species recovered from the splicing reaction shown inFigure 1(A) are presented following their immunoprecipitation with guinea anti-(72\74) (lanes 1-5) and…”

supporting

confidence: 76%

“…This type of experiment has been successfully applied in the past on β-globin pre-mRNA, using the well-known anti-Sm and anti-U1RNP antibodies, and has provided important information on the sites of binding of the spliceosomal snRNP complexes to pre-mRNA during in itro RNA splicing [25]. In our studies, we performed a similar type of analysis using the anti-(72\74) serum in parallel with a standard anti-Sm antibody [28]. The t l 15 min splicing reaction was selected as the time point for analysis, since this represents an early reaction time that combines the absence of intermediate products of splicing ( Figure 1A, lane 3) with the relatively high level of immunoselected β-globin pre-mRNA ( Figure 1B, lane 3).…”

Section: Specificity Of Binding Of the 72/74(m) Proteins To Pre-mrnamentioning

confidence: 99%

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Association of the 72/74-kDa proteins, members of the heterogeneous nuclear ribonucleoprotein M group, with the pre-mRNA at early stages of spliceosome assembly

Kafasla

Patrinou-Georgoula

Lewis

et al. 2002

Biochemical Journal

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We have investigated the role played in precursor mRNA (pre-mRNA) splicing by the protein pair of molecular size 72/74kDa, which are integral components of a discrete subset of heterogeneous nuclear (hn) ribonucleoproteins (RNPs) named large heterogeneous nuclear RNP (LH-nRNP). This 72/74kDa pair of proteins has been shown to belong to the hnRNP M group, and are referred to as 72/74(M). By applying specific immunoprecipitation assays in a consecutive series of splicing reactions in vitro, the antigenic 72/74(M) protein species were found to associate with the pre-mRNA and not the intermediate or final products of splicing. Kinetic studies, combined with isolation of pre-spliceosomal and spliceosomal complexes from the splicing reaction, indicated a loose association of 72/74(M) with both the initially formed H assembly and the first splicing-committed E complex. Stable binding was seen at a later stage of the reaction, well in advance of the appearance of the first intermediate products of RNA splicing. Evidence is provided that supports the almost exclusive association of 72/74(M) with pre-mRNA within the pre-spliceosomal A complex. This dynamic binding appeared to involve pre-mRNA sites similar to those of spliceosomal U1 and U2 small nuclear RNP complexes. Moreover, a preferential binding to a truncated RNA containing the 5′ exon—intron part, rather than the intron—3′ exon part, of pre-mRNA was observed.

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supporting

confidence: 76%

Section: Specificity Of Binding Of the 72/74(m) Proteins To Pre-mrnamentioning

confidence: 99%

Association of the 72/74-kDa proteins, members of the heterogeneous nuclear ribonucleoprotein M group, with the pre-mRNA at early stages of spliceosome assembly

Kafasla

Patrinou-Georgoula

Lewis

et al. 2002

Biochemical Journal

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“…Rat liver 40S hnRNP complexes were obtained after fractionation of nuclear extracts in 15-30% (w/v) sucrose gradients and subsequent pelleting of the material sedimenting in the 40-50S region of the gradient as described (25).…”

Section: Preparation Of 40s Pre-mrnp Fractionmentioning

confidence: 99%

Nuclear actin is associated with a specific subset of hnRNP A/B-type proteins

Percipalle

Jönsson

Nashchekin

et al. 2002

Nucleic Acids Research

144

112

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Pre-mRNP complexes were isolated from rat liver nuclei as 40S hnRNP particles, and actin-binding proteins were collected by DNase I affinity chromatography. The bound proteins were analyzed by 2D gel electrophoresis, and the following five hnRNP A/B-type proteins were identified by tandem mass spectrometry: DBP40/CBF-A (CArG binding factor A), a minor hnRNP A2 variant and three minor hnRNP A3 (mBx) variants. DBP40 was chosen for further analysis of the association of actin with the pre-mRNP complex. It was shown in vitro that purified actin binds to recombinant DBP40 suggesting that the interaction between actin and DBP40 is direct in the pre-mRNP particles. The association of actin with DBP40 was further explored in vivo. It was shown in a transfection study that DBP40 appears both in the nucleus and cytoplasm. Microinjection experiments revealed that DBP40 is exported from the nucleus to the cytoplasm. Finally, RNA-protein and protein-protein cross-linking experiments showed that DBP40 interacts with poly(A)(+) RNA as well as actin, both in the nucleus and cytoplasm. We propose that actin associated with DBP40, and perhaps with additional hnRNP A/B-type proteins, is transferred from nucleus to cytoplasm bound to mRNA.

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A fraction of the transcription factor TAF15 participates in interactions with a subset of the spliceosomal U1 snRNP complex

Leichter¹,

Marko²,

Ganou³

et al. 2011

Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics

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Structural/functional properties of a mammalian multi-component structure containing all major spliceosomal small nuclear ribonucleoprotein particles

Cited by 4 publications

References 37 publications

Association of the 72/74-kDa proteins, members of the heterogeneous nuclear ribonucleoprotein M group, with the pre-mRNA at early stages of spliceosome assembly

Association of the 72/74-kDa proteins, members of the heterogeneous nuclear ribonucleoprotein M group, with the pre-mRNA at early stages of spliceosome assembly

Nuclear actin is associated with a specific subset of hnRNP A/B-type proteins

A fraction of the transcription factor TAF15 participates in interactions with a subset of the spliceosomal U1 snRNP complex

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