Cancer-Associated Substitutions in RNA Recognition Motifs of
PUF60 and U2AF65 Reveal Residues Required for Correct Folding
and 3 ' Splice-Site Selection

J 2020

Cancer-Associated Substitutions in RNA Recognition Motifs of PUF60 and U2AF65 Reveal Residues Required for Correct Folding and 3 ' Splice-Site Selection

KRALOVICOVA, J., I. BOROVSKA, Monika KUBÍČKOVÁ, Peter LUKAVSKY, I. VORECHOVSKY et. al.

Základní údaje

Originální název

Cancer-Associated Substitutions in RNA Recognition Motifs of PUF60 and U2AF65 Reveal Residues Required for Correct Folding and 3 ' Splice-Site Selection

Autoři

KRALOVICOVA, J., I. BOROVSKA, Monika KUBÍČKOVÁ (203 Česká republika, domácí), Peter LUKAVSKY (40 Rakousko, garant, domácí) a I. VORECHOVSKY

Vydání

Cancers, BASEL, MDPI, 2020, 2072-6694

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Obor

30204 Oncology

Stát vydavatele

Švýcarsko

Utajení

není předmětem státního či obchodního tajemství

Odkazy

URL

Impakt faktor

Impact factor: 6.639

Kód RIV

RIV/00216224:14740/20:00118245

Organizační jednotka

Středoevropský technologický institut

DOI

http://dx.doi.org/10.3390/cancers12071865

UT WoS

000556370000001

Klíčová slova anglicky

Functional genomics; pre-mRNA splicing; 3 ' splice site; mRNA; lariat branch point; PUF60; U2AF2; gel shift assay; differential scanning fluorimetry; cancer; leukemia; driver mutation; exon inclusion

Štítky

rivok

Příznaky

Mezinárodní význam, Recenzováno

Změněno: 24. 2. 2021 10:30, Mgr. Pavla Foltynová, Ph.D.

Anotace

V originále

U2AF65 (U2AF2) and PUF60 (PUF60) are splicing factors important for recruitment of the U2 small nuclear ribonucleoprotein to lariat branch points and selection of 3 ' splice sites (3 ' ss). Both proteins preferentially bind uridine-rich sequences upstream of 3 ' ss via their RNA recognition motifs (RRMs). Here, we examined 36 RRM substitutions reported in cancer patients to identify variants that alter 3 ' ss selection, RNA binding and protein properties. Employing PUF60- and U2AF65-dependent 3 ' ss previously identified by RNA-seq of depleted cells, we found that 43% (10/23) and 15% (2/13) of independent RRM mutations in U2AF65 and PUF60, respectively, conferred splicing defects. At least three RRM mutations increased skipping of internalU2AF2(similar to 9%, 2/23) orPUF60(similar to 8%, 1/13) exons, indicating that cancer-associated RRM mutations can have bothcis- andtrans-acting effects on splicing. We also report residues required for correct folding/stability of each protein and map functional RRM substitutions on to existing high-resolution structures of U2AF65 and PUF60. These results identify new RRM residues critical for 3 ' ss selection and provide relatively simple tools to detect clonal RRM mutations that enhance the mRNA isoform diversity.

Návaznosti

630758, interní kód MU

Název: Aberrant Splicing of CFTR Exon 9 (Akronym: ASCE)

Investor: Evropská unie, Aberrant Splicing of CFTR Exon 9, Lidé

Citovat

KRALOVICOVA, J., I. BOROVSKA, Monika KUBÍČKOVÁ, Peter LUKAVSKY a I. VORECHOVSKY. Cancer-Associated Substitutions in RNA Recognition Motifs of PUF60 and U2AF65 Reveal Residues Required for Correct Folding and 3 ' Splice-Site Selection. Cancers. BASEL: MDPI, 2020, roč. 12, č. 7, s. 1865-1883. ISSN 2072-6694. Dostupné z: https://dx.doi.org/10.3390/cancers12071865.

@article{1745739,
   author = {Kralovicova, J. and Borovska, I. and Kubíčková, Monika and Lukavsky, Peter and Vorechovsky, I.},
   article_location = {BASEL},
   article_number = {7},
   doi = {http://dx.doi.org/10.3390/cancers12071865},
   keywords = {Functional genomics; pre-mRNA splicing; 3 ' splice site; mRNA; lariat branch point; PUF60; U2AF2; gel shift assay; differential scanning fluorimetry; cancer; leukemia; driver mutation; exon inclusion},
   language = {eng},
   issn = {2072-6694},
   journal = {Cancers},
   title = {Cancer-Associated Substitutions in RNA Recognition Motifs of PUF60 and U2AF65 Reveal Residues Required for Correct Folding and 3 ' Splice-Site Selection},
   url = {https://www.mdpi.com/2072-6694/12/7/1865},
   volume = {12},
   year = {2020}
}

TY  - JOUR
ID  - 1745739
AU  - Kralovicova, J. - Borovska, I. - Kubíčková, Monika - Lukavsky, Peter - Vorechovsky, I.
PY  - 2020
TI  - Cancer-Associated Substitutions in RNA Recognition Motifs of PUF60 and U2AF65 Reveal Residues Required for Correct Folding and 3 ' Splice-Site Selection
JF  - Cancers
VL  - 12
IS  - 7
SP  - 1865
EP  - 1865
PB  - MDPI
SN  - 20726694
KW  - Functional genomics
KW  - pre-mRNA splicing
KW  - 3 ' splice site
KW  - mRNA
KW  - lariat branch point
KW  - PUF60
KW  - U2AF2
KW  - gel shift assay
KW  - differential scanning fluorimetry
KW  - cancer
KW  - leukemia
KW  - driver mutation
KW  - exon inclusion
UR  - https://www.mdpi.com/2072-6694/12/7/1865
N2  - U2AF65 (U2AF2) and PUF60 (PUF60) are splicing factors important for recruitment of the U2 small nuclear ribonucleoprotein to lariat branch points and selection of 3 ' splice sites (3 ' ss). Both proteins preferentially bind uridine-rich sequences upstream of 3 ' ss via their RNA recognition motifs (RRMs). Here, we examined 36 RRM substitutions reported in cancer patients to identify variants that alter 3 ' ss selection, RNA binding and protein properties. Employing PUF60- and U2AF65-dependent 3 ' ss previously identified by RNA-seq of depleted cells, we found that 43% (10/23) and 15% (2/13) of independent RRM mutations in U2AF65 and PUF60, respectively, conferred splicing defects. At least three RRM mutations increased skipping of internalU2AF2(similar to 9%, 2/23) orPUF60(similar to 8%, 1/13) exons, indicating that cancer-associated RRM mutations can have bothcis- andtrans-acting effects on splicing. We also report residues required for correct folding/stability of each protein and map functional RRM substitutions on to existing high-resolution structures of U2AF65 and PUF60. These results identify new RRM residues critical for 3 ' ss selection and provide relatively simple tools to detect clonal RRM mutations that enhance the mRNA isoform diversity.
ER  -

KRALOVICOVA, J., I. BOROVSKA, Monika KUBÍČKOVÁ, Peter LUKAVSKY a I. VORECHOVSKY. Cancer-Associated Substitutions in RNA Recognition Motifs of PUF60 and U2AF65 Reveal Residues Required for Correct Folding and 3 ' Splice-Site Selection. \textit{Cancers}. BASEL: MDPI, 2020, roč.~12, č.~7, s.~1865-1883. ISSN~2072-6694. Dostupné z: https://dx.doi.org/10.3390/cancers12071865.

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