Central Region of the Human Splicing Factor Hprp3p Interacts with Hprp4p

Gonzalez-Santos, Juana Maria; Wang, Anan; Jones, Joses R. W.; Ushida, Chisato; Liu, Jun; Hu, Jim

doi:10.1074/jbc.m111461200

Cited by 29 publications

(43 citation statements)

References 47 publications

(53 reference statements)

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“…Regarding the function, hPrp3p is a new type of binding partner, because it does not belong to the class of proteins involved in cellular signalling, cell cycle control or intracellular trafficking, instead hPrp3p is implicated in splicing. Together with hPrp4p hPrp3p is associated with the U4/U6 small nuclear ribonucleoprotein particle, which is essential for the assembly of an active spliceosome (Gonzalez-Santos et al, 2002). These snRNAs recognize conserved sequences of the premRNA and assemble into a catalytically active spliceosome that catalyses the cleavage/ligation reactions of RNA splicing.…”

Section: Discussionmentioning

confidence: 99%

Protein kinase CK2 interacts with the splicing factor hPrp3p

et al. 2007

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Numerous signalling pathways in cells are influenced by the ubiquitous Ser/Thr protein kinase CK2. Protein kinase CK2 is composed of two regulatory b-subunits and two catalytic a-or a 0 -subunits. Several of the known CK2 substrates are proteins known to regulate transcriptional events. Here, we describe that protein kinase CK2 interacts with the splicing factor hPrp3p, which is important for the assembly of the spliceosome. In a twohybrid screen hPrp3p is exclusively bound to the catalytic a-or a 0 -subunits of CK2 but not to the regulatory bsubunit. The interaction was confirmed by coimmunoprecipitation experiments in vitro and in vivo. Moreover, both proteins colocalized in nuclear speckles which is typical for splicing factor compartments within the nucleus. Phosphorylation experiments revealed that hPrp3p is also a substrate of protein kinase CK2. The main phosphorylation site was mapped to C-terminal residues. In vitro and in vivo splicing assays showed that the splicing activity is significantly influenced by the CK2-hPrp3p interaction. Thus, these data showed that CK2 is involved in the regulation of RNA processing.

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Section: Discussionmentioning

confidence: 99%

Protein kinase CK2 interacts with the splicing factor hPrp3p

et al. 2007

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“…Like PRPF31p, HPRP3 is a component of the U4/U6 snRNP (Wang et al 1997). In mammals, HPRP3 is thought to recruit HPRP4 to the U4/U6 snRNP (Gonzalez-Santos et al 2002). HPRP3 and HPRP4 are homologs of the yeast U4/U6-snRNP-specific proteins.…”

Section: Retinitis Pigmentosamentioning

confidence: 99%

Pre-mRNA splicing and human disease

Faustino¹,

Cooper²

2003

Genes Dev.

1,127

862

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The precision and complexity of intron removal during pre-mRNA splicing still amazes even 26 years after the discovery that the coding information of metazoan genes is interrupted by introns (Berget et al. 1977;Chow et al. 1977). Adding to this amazement is the recent realization that most human genes express more than one mRNA by alternative splicing, a process by which functionally diverse protein isoforms can be expressed according to different regulatory programs. Given that the vast majority of human genes contain introns and that most pre-mRNAs undergo alternative splicing, it is not surprising that disruption of normal splicing patterns can cause or modify human disease. The purpose of this review is to highlight the different mechanisms by which disruption of pre-mRNA splicing play a role in human disease. Several excellent reviews provide detailed information on splicing and the regulation of splicing (Burge et al. 1999;Hastings and Krainer 2001; Black 2003). The potential role of splicing as a modifier of human disease has also recently been reviewed (NissimRafinia and Kerem 2002). Constitutive splicing and the basal splicing machineryThe typical human gene contains an average of 8 exons. Internal exons average 145 nucleotides (nt) in length, and introns average more than 10 times this size and can be much larger (Lander et al. 2001). Exons are defined by rather short and degenerate classical splice-site sequences at the intron/exon borders (5Ј splice site, 3Ј splice site, and branch site; Fig. 1A). Components of the basal splicing machinery bind to the classical splice-site sequences and promote assembly of the multicomponent splicing complex known as the spliceosome. The spliceosome performs the two primary functions of splicing: recognition of the intron/exon boundaries and catalysis of the cut-and-paste reactions that remove introns and join exons. The spliceosome is made up of five small nuclear ribonucleoproteins (snRNPs) and >100 proteins. Each snRNP is composed of a single uridinerich small nuclear RNA (snRNA) and multiple proteins. The U1 snRNP binds the 5Ј splice site, and the U2 snRNP binds the branch site via RNA:RNA interactions between the snRNA and the pre-mRNA (Fig. 1B). Spliceosome assembly is highly dynamic in that complex rearrangements of RNA:RNA, RNA:protein, and protein:protein interactions take place within the spliceosome. Coinciding with these internal rearrangements, both splice sites are recognized multiple times by interactions with different components during the course of spliceosome assembly (for example, see Burge et al. 1999;Du and Rosbash 2002;Lallena et al. 2002;Liu 2002). The catalytic component is likely to be U6 snRNP, which joins the spliceosome as a U4/U6 · U5 tri-snRNP (Villa et al. 2002).A splicing error that adds or removes even 1 nt will disrupt the open reading frame of an mRNA; yet exons are correctly spliced from within tens of thousands of intronic nucleotides. This remarkable precision is, in part, built into the mechanism of intron removal because once...

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“…Notably, no interaction was found between the 15.5K protein and any of the other proteins. As binding between hPrp4 and the central domain of hPrp3 had previously been demonstrated using co-immunoprecipitation and isothermal titration calorimetry (Gonzalez-Santos et al 2002), we also performed in vitro binding assays with Histagged hPrp3 protein. Significantly, in vitro translated HAtagged hPrp4 was co-precipitated specifically and efficiently by His-tagged hPrp3, confirming this interaction (not shown).…”

Section: Analysis Of Interactions Between U4/u6 Proteinsmentioning

confidence: 99%

“…1). To delineate the boundaries of the hSnu66-interacting domain, hPrp3 was divided into four fragments according to (Gonzalez-Santos et al 2002) and two additional fragments comprising the N-and C-terminal halves of the protein (Fig. 6A).…”

Section: Reassessment Of Y2h Positives In the Presence Of 3-atmentioning

confidence: 99%

“…In contrast, relatively little is known about the proteinprotein and protein-RNA network within the human trisnRNP. The human U4/U6-specific proteins CypH, hPrp4, and hPrp3 form a stable RNA-free trimeric subcomplex, and hPrp4 and hPrp3 have been shown to interact directly with each other (Horowitz et al 1997;Lauber et al 1997;Teigelkamp et al 1998;Gonzalez-Santos et al 2002). An additional stable RNA-free subcomplex is formed by the U5 proteins hSnu114, hPrp8, hBrr2, and 40K .…”

Section: Introductionmentioning

confidence: 99%

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The network of protein–protein interactions within the human U4/U6.U5 tri-snRNP

Liu¹,

Rauhut²,

Vornlocher³

et al. 2006

RNA

172

208

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The human 25S U4/U6.U5 tri-snRNP is a major building block of the U2-type spliceosome and contains, in addition to the U4, U6, and U5 snRNAs, at least 30 distinct proteins. To learn more about the molecular architecture of the tri-snRNP, we have investigated interactions between tri-snRNP proteins using the yeast two-hybrid assay and in vitro binding assays, and, in addition, have identified distinct protein domains that are critical for the connectivity of this protein network in the human tri-snRNP. These studies revealed multiple interactions between distinct domains of the U5 proteins hPrp8, hBrr2 (a DExH/Dbox helicase), and hSnu114 (a putative GTPase), which are key players in the catalytic activation of the spliceosome, during which the U4/U6 base-pairing interaction is disrupted and U4 is released from the spliceosome. Both the U5-specific, TPR/HATrepeat-containing hPrp6 protein and the tri-snRNP-specific hSnu66 protein interact with several U5-and U4/U6-associated proteins, including hBrr2 and hPrp3, which contacts the U6 snRNA. Thus, both proteins are located at the interface between U5 and U4/U6 in the tri-snRNP complex, and likely play an important role in transmitting the activity of hBrr2 and hSnu114 in the U5 snRNP to the U4/U6 duplex during spliceosome activation. A more detailed analysis of these protein interactions revealed that different HAT repeats mediate interactions with specific hPrp6 partners. Taken together, data presented here provide a detailed picture of the network of protein interactions within the human tri-snRNP.

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Central Region of the Human Splicing Factor Hprp3p Interacts with Hprp4p

Cited by 29 publications

References 47 publications

Protein kinase CK2 interacts with the splicing factor hPrp3p

Protein kinase CK2 interacts with the splicing factor hPrp3p

Pre-mRNA splicing and human disease

The network of protein–protein interactions within the human U4/U6.U5 tri-snRNP

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