Magnesium Cations are Required for the Association of U Small Nuclear Ribonucleoproteins and SR Proteins with Pre-mRNA in 200 S Large Nuclear Ribonucleoprotein Particles

Miriami, Elana; Angenitzki, Mina; Sperling, Ruth; Sperling, Joseph

doi:10.1006/jmbi.1994.0081

Cited by 28 publications

(60 citation statements)

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“…The heaviest fraction pelleted to the bottom of the gradient and did so even when centrifugation speeds and times were significantly shortened, suggesting that these complexes represent non-specific aggregates which may be similar to previously described complexes. [9][10][11] However, fractions 6-11 contained PSF in complexes similar in size to mammalian spliceosomes (as indicated by the peak of ADML pre-mRNA, mRNA and splicing intermediates in parallel gradients, Fig. 2B, fractions 6-9).…”

Section: Resultsmentioning

confidence: 89%

“…In recent years, several complexes have been identified that seem to challenge previous models of spliceosome formation. These include the yeast Cwf/Cwc complex, 7,8 lnRNP complexes, 9 a complex larger than 150S with base paired U2 and U6, 10 a U2/U4/U5/U6-containing pseudospliceosome complex, 11 a pentasnRNP from S. cerevisiae, 12 and a HeLa nuclear extract penta-snRNP complex. 13 The exact functional relevance of these complexes remains to be determined, none have been shown to be capable of splicing on their own.…”

Section: Introductionmentioning

confidence: 99%

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The Splicing Factor PSF is Part of a Large Complex That Assembles in the Absence of pre-mRNA and Contains All 5 snRNPs

Peng¹,

Hawkins²,

Link³

et al. 2006

RNA Biology

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Previously published online as a RNA Biology E-publication: http://www.landesbioscience.com/journals/rnabiology/abstract.php?id=3017 KEY WORDS Research PaperThe Splicing Factor PSF is Part of a Large Complex that Assembles in the Absence of pre-mRNA and Contains All 5 snRNPs ABSTRACT PSF (PTB-associated splicing factor) is a large nuclear protein that has been implicated in numerous processes including transcription and RNA splicing. It has been shown to directly associate with U5 snRNA and has also been found within numerous purified splicing complexes. Here, we show that when HeLa nuclear extracts are adjusted to splicing conditions, PSF is found as part of a large complex that contains all five snRNPs and most known splicing factors. Formation of the complex does not require addition of exogenous pre-mRNA substrate and occurs at 4˚C but is salt sensitive. Sedimentation experiments and identification of individual components by mass spectrometry revealed association with multiple nuclear factors, most of which overlap with spliceosome components.

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Section: Resultsmentioning

confidence: 89%

Section: Introductionmentioning

confidence: 99%

The Splicing Factor PSF is Part of a Large Complex That Assembles in the Absence of pre-mRNA and Contains All 5 snRNPs

Peng¹,

Hawkins²,

Link³

et al. 2006

RNA Biology

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“…These processes have been proposed to be carried out within large nuclear ribonucleoprotein (lnRNP) particles that sediment at 200S in sucrose gradients. The composition of the lnRNP particles (19)(20)(21)(22) is very similar to that of the in vitro-assembled spliceosome (reviewed in refs. 23 and 24).…”

mentioning

confidence: 94%

“…23 and 24). Specifically, all of the U small nuclear ribonucleoproteins (snRNPs) required for pre-mRNA splicing, as well as several non-snRNP protein splicing factors, including U2AF and the SR protein family (23)(24)(25) are present within the lnRNP particles (19,21,22).…”

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confidence: 99%

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RNA editing activity is associated with splicing factors in lnRNP particles: The nuclear pre-mRNA processing machinery

Raitskin

Cho

Sperling

et al. 2001

Proc. Natl. Acad. Sci. U.S.A.

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Multiple members of the ADAR (adenosine deaminases acting on RNA) gene family are involved in A-to-I RNA editing. It has been speculated that they may form a large multicomponent protein complex. Possible candidates for such complexes are large nuclear ribonucleoprotein (lnRNP) particles. The lnRNP particles consist mainly of four spliceosomal subunits that assemble together with the pre-mRNA to form a large particle and thus are viewed as the naturally assembled pre-mRNA processing machinery. Here we investigated the presence of ADARs in lnRNP particles by Western blot analysis using anti-ADAR antibodies and by indirect immunoprecipitation. Both ADAR1 and ADAR2 were found associated with the spliceosomal components Sm and SR proteins within the lnRNP particles. The two ADARs, associated with lnRNP particles, were enzymatically active in site-selective A-to-I RNA editing. We demonstrate the association of ADAR RNA editing enzymes with physiological supramolecular complexes, the lnRNP particles.adenosine deaminases acting on RNA ͉ double-stranded RNA adenosine deaminase ͉ nucleic acid-protein interactions ͉ large nuclear ribonucleoprotein

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Ribonucleoprotein ( RNP )

Sperling¹,

Sperling²

2002

Encyclopedia of Molecular Biology

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Magnesium Cations are Required for the Association of U Small Nuclear Ribonucleoproteins and SR Proteins with Pre-mRNA in 200 S Large Nuclear Ribonucleoprotein Particles

Cited by 28 publications

References 0 publications

The Splicing Factor PSF is Part of a Large Complex That Assembles in the Absence of pre-mRNA and Contains All 5 snRNPs

The Splicing Factor PSF is Part of a Large Complex That Assembles in the Absence of pre-mRNA and Contains All 5 snRNPs

RNA editing activity is associated with splicing factors in lnRNP particles: The nuclear pre-mRNA processing machinery

Ribonucleoprotein ( RNP )

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