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ZNF265--a novel spliceosomal protein able to induce alternative splicing.

Adams DJ, van der Weyden L, Mayeda A, Stamm S, Morris BJ, Rasko JE - J. Cell Biol. (2001)

Bottom Line: Transfection of HT-1080 cells with ZNF265-EGFP fusion constructs showed that nuclear localization of ZNF265 required the RS domain.Alignment with other RS domain-containing proteins revealed a high degree of SR dipeptide conservation.These data show that ZNF265 functions as a novel component of the mRNA processing machinery.

View Article: PubMed Central - PubMed

Affiliation: The University of Sydney, Basic & Clinical Genomics Laboratory, Department of Physiology and Institute for Biomedical Research, Sydney, NSW 2006, Australia.

ABSTRACT
The formation of the active spliceosome, its recruitment to active areas of transcription, and its role in pre-mRNA splicing depends on the association of a number of multifunctional serine/arginine-rich (SR) proteins. ZNF265 is an arginine/serine-rich (RS) domain containing zinc finger protein with conserved pre-mRNA splicing protein motifs. Here we show that ZNF265 immunoprecipitates from splicing extracts in association with mRNA, and that it is able to alter splicing patterns of Tra2-beta1 transcripts in a dose-dependent manner in HEK 293 cells. Yeast two-hybrid analysis and immunoprecipitation indicated interaction of ZNF265 with the essential splicing factor proteins U1-70K and U2AF(35). Confocal microscopy demonstrated colocalization of ZNF265 with the motor neuron gene product SMN, the snRNP protein U1-70K, the SR protein SC35, and with the transcriptosomal components p300 and YY1. Transfection of HT-1080 cells with ZNF265-EGFP fusion constructs showed that nuclear localization of ZNF265 required the RS domain. Alignment with other RS domain-containing proteins revealed a high degree of SR dipeptide conservation. These data show that ZNF265 functions as a novel component of the mRNA processing machinery.

Show MeSH
Conservation of serine and arginine residues in ZNF265 and other RS domain proteins. Alignment of the RS domain of ZNF265 (NP_005446) with RS domains of the spliceosomal proteins SF2/ASF (NP_008855), SC35 (A42634), SRp20 (NP_003008), SRp40 (S59042), SRp55 (S59043), SRp75 (A48133), U1-70K (A25707), U2AF35 (Q01081), U2AF65 (NP_009210), 9G8 (A57198), and p54 (XP_001835). Sequence alignment was performed using “Pileup” and “Prettybox” (Australian Genomic Information Service). Residues conserved in the majority of the aligned proteins are shaded. Numbers indicate the amino acid position of each protein.
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fig4: Conservation of serine and arginine residues in ZNF265 and other RS domain proteins. Alignment of the RS domain of ZNF265 (NP_005446) with RS domains of the spliceosomal proteins SF2/ASF (NP_008855), SC35 (A42634), SRp20 (NP_003008), SRp40 (S59042), SRp55 (S59043), SRp75 (A48133), U1-70K (A25707), U2AF35 (Q01081), U2AF65 (NP_009210), 9G8 (A57198), and p54 (XP_001835). Sequence alignment was performed using “Pileup” and “Prettybox” (Australian Genomic Information Service). Residues conserved in the majority of the aligned proteins are shaded. Numbers indicate the amino acid position of each protein.

Mentions: Alignment of the RS domain of ZNF265 with that of other RS domain–containing spliceosomal proteins (Fig. 4) showed strong SR dipeptide conservation; this was particularly evident between ZNF265, SC35, and SRp40. The aligned region of SC35 contains the putative RS domain NLS, RRRRRSRSRSRSRSRSRSRSRYSRSKSRSR-TRSRSRSTSKSRS (Hedley et al., 1995).


ZNF265--a novel spliceosomal protein able to induce alternative splicing.

Adams DJ, van der Weyden L, Mayeda A, Stamm S, Morris BJ, Rasko JE - J. Cell Biol. (2001)

Conservation of serine and arginine residues in ZNF265 and other RS domain proteins. Alignment of the RS domain of ZNF265 (NP_005446) with RS domains of the spliceosomal proteins SF2/ASF (NP_008855), SC35 (A42634), SRp20 (NP_003008), SRp40 (S59042), SRp55 (S59043), SRp75 (A48133), U1-70K (A25707), U2AF35 (Q01081), U2AF65 (NP_009210), 9G8 (A57198), and p54 (XP_001835). Sequence alignment was performed using “Pileup” and “Prettybox” (Australian Genomic Information Service). Residues conserved in the majority of the aligned proteins are shaded. Numbers indicate the amino acid position of each protein.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2196870&req=5

fig4: Conservation of serine and arginine residues in ZNF265 and other RS domain proteins. Alignment of the RS domain of ZNF265 (NP_005446) with RS domains of the spliceosomal proteins SF2/ASF (NP_008855), SC35 (A42634), SRp20 (NP_003008), SRp40 (S59042), SRp55 (S59043), SRp75 (A48133), U1-70K (A25707), U2AF35 (Q01081), U2AF65 (NP_009210), 9G8 (A57198), and p54 (XP_001835). Sequence alignment was performed using “Pileup” and “Prettybox” (Australian Genomic Information Service). Residues conserved in the majority of the aligned proteins are shaded. Numbers indicate the amino acid position of each protein.
Mentions: Alignment of the RS domain of ZNF265 with that of other RS domain–containing spliceosomal proteins (Fig. 4) showed strong SR dipeptide conservation; this was particularly evident between ZNF265, SC35, and SRp40. The aligned region of SC35 contains the putative RS domain NLS, RRRRRSRSRSRSRSRSRSRSRYSRSKSRSR-TRSRSRSTSKSRS (Hedley et al., 1995).

Bottom Line: Transfection of HT-1080 cells with ZNF265-EGFP fusion constructs showed that nuclear localization of ZNF265 required the RS domain.Alignment with other RS domain-containing proteins revealed a high degree of SR dipeptide conservation.These data show that ZNF265 functions as a novel component of the mRNA processing machinery.

View Article: PubMed Central - PubMed

Affiliation: The University of Sydney, Basic & Clinical Genomics Laboratory, Department of Physiology and Institute for Biomedical Research, Sydney, NSW 2006, Australia.

ABSTRACT
The formation of the active spliceosome, its recruitment to active areas of transcription, and its role in pre-mRNA splicing depends on the association of a number of multifunctional serine/arginine-rich (SR) proteins. ZNF265 is an arginine/serine-rich (RS) domain containing zinc finger protein with conserved pre-mRNA splicing protein motifs. Here we show that ZNF265 immunoprecipitates from splicing extracts in association with mRNA, and that it is able to alter splicing patterns of Tra2-beta1 transcripts in a dose-dependent manner in HEK 293 cells. Yeast two-hybrid analysis and immunoprecipitation indicated interaction of ZNF265 with the essential splicing factor proteins U1-70K and U2AF(35). Confocal microscopy demonstrated colocalization of ZNF265 with the motor neuron gene product SMN, the snRNP protein U1-70K, the SR protein SC35, and with the transcriptosomal components p300 and YY1. Transfection of HT-1080 cells with ZNF265-EGFP fusion constructs showed that nuclear localization of ZNF265 required the RS domain. Alignment with other RS domain-containing proteins revealed a high degree of SR dipeptide conservation. These data show that ZNF265 functions as a novel component of the mRNA processing machinery.

Show MeSH