The 5′ terminus of the RNA moiety of U1 small nuclear ribonucleoprotein particles is required for the splicing of messenger RNA precursors

Krämer, Angela; Keller, Walter; Appel, Bernd; Lührmann, Reinhard

doi:10.1016/0092-8674(84)90551-8

Cited by 407 publications

(230 citation statements)

References 29 publications

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“…Thus, the existence of a histone mRNA 3' processing activity in Xenopus laevis oocytes and Drosophila melanogaster tissue culture cell extracts has been clearly demonstrated (4,25,43). Furthermore, it has been shown that this activity is associated with a small nuclear RNA U7 (12, 52) in a similar way to the involvement of the small nuclear RNA Ul in intronic splicing (24,27,42,47). These results indicate that transcriptional termination is likely to occur separately from 3'-end processing in histone genes.…”

mentioning

confidence: 73%

Tripartite sequences within and 3' to the sea urchin H2A histone gene display properties associated with a transcriptional termination process.

Johnson¹,

Norman²,

Reeve³

et al. 1986

Mol. Cell. Biol.

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We have defined a DNA sequence that behaves as an RNA polymerase II termination signal by using the human HeLa cell transient expression system. Surprisingly, this sequence is tripartite, including part of the coding region of the sea urchin H2A histone gene together with two separate sequences in the 3' flanking region of the gene. We demonstrate that this signal functions both in its normal gene environment and also when placed within the human a-globin gene. However, we have failed to detect a discrete 3' terminus. Rather, our data indicate the, presence of an extremely heterogeneous series of nonpolyadenylated RNAs. These heterogeneous nonpolyadenylated RNAs are stable when transcribed from the intact histone gene but are highly unstable within the human a-globin gene. This provides evidence for the role of poly(A) in the stability of mnRNA.Initiation of transcription in eucaryotic RNA polymerase II (polII) genes is a highly regulated and in some instances well-defined process (see reference 28 for a review). In contrast, termination of transcription in polII genes is ill defined and may not be a regulated process. The principal cause of this uncertainty is the rapid posttranscriptional processing of primary RNA transcripts to mature mRNA (see reference 40 for a review). The fact that introns are spliced out of the polII gene transcript is well documented. Similarly, the generation of mRNA 3' termini for both polyadenylated [poly(A)+] mRNA and nonpolyadenylated [poly(A)-] histone mRNA by a processing event has been recently established (see references 5 and 44 for reviews).The best evidence for processing of mRNA 3' ends comes from studies on histone mRNA. Thus, the existence of a histone mRNA 3' processing activity in Xenopus laevis oocytes and Drosophila melanogaster tissue culture cell extracts has been clearly demonstrated (4,25,43). Furthermore, it has been shown that this activity is associated with a small nuclear RNA U7 (12, 52) in a similar way to the involvement of the small nuclear RNA Ul in intronic splicing (24,27,42,47). These results indicate that transcriptional termination is likely to occur separately from 3'-end processing in histone genes. Evidence of a similar nature in poly(A)+ mRNA genes is now becoming available. Moore and Sharp (37) have recently shown that synthetic RNAs extending beyond an adenovirus poly(A) site can be efficiently and accurately cleaved and polyadenylated by using in vitro cell extracts.Further evidence for transcriptional termination separate from nmRNA 3'-end formation comes from hybridization analysis of pulse-labeled RNA transcripts. Using these techniques, Nevins and Darnell (38) and Fraser et al. (11) deduced that the major late gene transcripts of adenovirus extend beyond the various late mRNA poly(A) sites to near the end of the viral genome. Similarly, transcription extends beyond the adenovirus early region 2 and 4 poly(A) sites and the simian virus 40 (SV40) late poly(A) site (10, 39). These viral transcription experiments were performed on pulse-* Corr...

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confidence: 73%

Tripartite sequences within and 3' to the sea urchin H2A histone gene display properties associated with a transcriptional termination process.

Johnson¹,

Norman²,

Reeve³

et al. 1986

Mol. Cell. Biol.

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“…7he in vitro 3' processing reaction of mouse H4 pre-mRNA is dependent on small nuclear RNPs We wished to investigate whether snRNPs are involved in the in vitro 3' processing reaction as this was shown to be the case for the maturation of sea urchin H3 pre-mRNA in frog oocyte injection experiments (Galli et al, 1983). For this, aliquots of the nuclear extracts were preincubated in the presence and in the absence of Sm-antiserum (Kramer et al, 1984) which is known to remove U-snRNPs. The extracts treated in this way, as well as extracts incubated with control sera and non-incubated extracts were challenged with the SP64-H4-257/63 precursor RNA.…”

Section: Resultsmentioning

confidence: 99%

Generation of histone mRNA 3′ ends by endonucleolytic cleavage of the pre-mRNA in a snRNP-dependent in vitro reaction.

Gick¹,

Krämer²,

Keller³

et al. 1986

The EMBO Journal

146

135

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Incubation of SP6 generated mouse histone H4 mRNA precursors in nuclear extracts of HeLa cells yields processed mRNA species which end on the 3' adenosine of the conserved terminal ACCA sequence not unlike ten different histone mRNAs isolated from sea urchin embryos which end either on the 3' C or A. In the presence of 20 mM EDTA, the cleaved off 3' spacer portions of the RNA transcripts are readily detected, hence the 3' ends of histone mRNAs arise as a consequence of endonucleolytic cleavage(s) of the precursor RNA. The in vitro cleavage reaction is specifically inhibited by human antisera of the Sm-serotype, but not by control sera and can be rescued by the addition of a preparation of partially purified small nuclear RNPs to the antibody-depleted extract. Interestingly, the snRNP preparation is sufficient to elicit 3' processing of pre-mRNA in the absence of added nuclear extract.

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“…The sequence of this RNA has been highly conserved in evolution, with only a 25 to 30% difference between Ul RNA from such diverse organisms as rats, drosophila, and sea urchins (3,5,28). Much evidence is accumulating that Ul RNA functions in the biosynthesis of mRNA at the level of RNA splicing (17,27,31). However, little is known about the biosynthesis of Ul RNA.…”

mentioning

confidence: 99%

Synthesis of U1 RNA in isolated nuclei from sea urchin embryos: U1 RNA is initiated at the first nucleotide of the RNA.

Morris¹,

Marzluff²

1985

Mol. Cell. Biol.

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Nuclei from sea urchin blastula embryos synthesize a variety of small RNAs, one of which has identical mobility with sea urchin Ul RNA. This RNA is synthesized by RNA polymerase II and, in a hybridizationselection experiment, was selected by the cloned sea urchin Ul gene. The Ul RNA was initiated with ATP, but not GIP, in isolated nuclei with P-S-and y-S-ribonucleotide triphosphates as substrates. The Ul RNA containing thiophosphate at the 5' end was not capped but accumulated as an uncapped transcript from which the thiophosphate could be removed with calf intestinal phosphatase.

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The 5′ terminus of the RNA moiety of U1 small nuclear ribonucleoprotein particles is required for the splicing of messenger RNA precursors

Cited by 407 publications

References 29 publications

Tripartite sequences within and 3' to the sea urchin H2A histone gene display properties associated with a transcriptional termination process.

Tripartite sequences within and 3' to the sea urchin H2A histone gene display properties associated with a transcriptional termination process.

Generation of histone mRNA 3′ ends by endonucleolytic cleavage of the pre-mRNA in a snRNP-dependent in vitro reaction.

Synthesis of U1 RNA in isolated nuclei from sea urchin embryos: U1 RNA is initiated at the first nucleotide of the RNA.

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