PMID- 33317029
OWN - NLM
STAT- MEDLINE
DCOM- 20210621
LR  - 20210621
IS  - 2073-4409 (Electronic)
IS  - 2073-4409 (Linking)
VI  - 9
IP  - 12
DP  - 2020 Dec 9
TI  - U2AF65-Dependent SF3B1 Function in SMN Alternative Splicing.
LID - 10.3390/cells9122647 [doi]
LID - 2647
AB  - Splicing factor 3b subunit 1 (SF3B1) is an essential protein in spliceosomes and 
      mutated frequently in many cancers. While roles of SF3B1 in single intron 
      splicing and roles of its cancer-linked mutant in aberrant splicing have been 
      identified to some extent, regulatory functions of wild-type SF3B1 in alternative 
      splicing (AS) are not well-understood yet. Here, we applied RNA sequencing 
      (RNA-seq) to analyze genome-wide AS in SF3B1 knockdown (KD) cells and to identify 
      a large number of skipped exons (SEs), with a considerable number of alternative 
      5' splice-site selection, alternative 3' splice-site selection, mutually 
      exclusive exons (MXE), and retention of introns (RI). Among altered SEs by SF3B1 
      KD, survival motor neuron 2 (SMN2) pre-mRNA exon 7 splicing was a regulatory 
      target of SF3B1. RT-PCR analysis of SMN exon 7 splicing in SF3B1 KD or 
      overexpressed HCT116, SH-SY5Y, HEK293T, and spinal muscular atrophy (SMA) patient 
      cells validated the results. A deletion mutation demonstrated that the U2 snRNP 
      auxiliary factor 65 kDa (U2AF65) interaction domain of SF3B1 was required for its 
      function in SMN exon 7 splicing. In addition, mutations to lower the score of the 
      polypyrimidine tract (PPT) of exon 7, resulting in lower affinity for U2AF65, 
      were not able to support SF3B1 function, suggesting the importance of U2AF65 in 
      SF3B1 function. Furthermore, the PPT of exon 7 with higher affinity to U2AF65 
      than exon 8 showed significantly stronger interactions with SF3B1. Collectively, 
      our results revealed SF3B1 function in SMN alternative splicing.
FAU - Choi, Namjeong
AU  - Choi N
AD  - School of Life Sciences, Gwangju Institute of Science and Technology, Gwangju 
      500-712, Korea.
FAU - Liu, Yongchao
AU  - Liu Y
AD  - School of Life Sciences, Gwangju Institute of Science and Technology, Gwangju 
      500-712, Korea.
FAU - Oh, Jagyeong
AU  - Oh J
AD  - School of Life Sciences, Gwangju Institute of Science and Technology, Gwangju 
      500-712, Korea.
FAU - Ha, Jiyeon
AU  - Ha J
AD  - School of Life Sciences, Gwangju Institute of Science and Technology, Gwangju 
      500-712, Korea.
FAU - Zheng, Xuexiu
AU  - Zheng X
AD  - School of Life Sciences, Gwangju Institute of Science and Technology, Gwangju 
      500-712, Korea.
FAU - Shen, Haihong
AU  - Shen H
AD  - School of Life Sciences, Gwangju Institute of Science and Technology, Gwangju 
      500-712, Korea.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20201209
PL  - Switzerland
TA  - Cells
JT  - Cells
JID - 101600052
RN  - 0 (Phosphoproteins)
RN  - 0 (RNA Precursors)
RN  - 0 (RNA Splicing Factors)
RN  - 0 (RNA, Small Interfering)
RN  - 0 (SF3B1 protein, human)
RN  - 0 (SMN2 protein, mouse)
RN  - 0 (Smn1 protein, mouse)
RN  - 0 (Splicing Factor U2AF)
RN  - 0 (Survival of Motor Neuron 1 Protein)
RN  - 0 (Survival of Motor Neuron 2 Protein)
RN  - 139076-35-0 (Polypyrimidine Tract-Binding Protein)
SB  - IM
MH  - *Alternative Splicing
MH  - Cell Line
MH  - Exons
MH  - Humans
MH  - Muscular Atrophy, Spinal/metabolism/pathology
MH  - Phosphoproteins/antagonists & inhibitors/genetics/*metabolism
MH  - Polypyrimidine Tract-Binding Protein/genetics/metabolism
MH  - Protein Binding
MH  - RNA Interference
MH  - RNA Precursors/genetics/metabolism
MH  - RNA Splicing Factors/antagonists & inhibitors/genetics/*metabolism
MH  - RNA, Small Interfering/metabolism
MH  - Splicing Factor U2AF/chemistry/*metabolism
MH  - Survival of Motor Neuron 1 Protein/*genetics/metabolism
MH  - Survival of Motor Neuron 2 Protein/*genetics/metabolism
PMC - PMC7762998
OTO - NOTNLM
OT  - SF3B1
OT  - SMN
OT  - alternative splicing
OT  - spinal muscular atrophy
COIS- The authors declare no conflict of interest.
EDAT- 2020/12/16 06:00
MHDA- 2021/06/22 06:00
PMCR- 2020/12/09
CRDT- 2020/12/15 01:04
PHST- 2020/11/14 00:00 [received]
PHST- 2020/12/05 00:00 [revised]
PHST- 2020/12/07 00:00 [accepted]
PHST- 2020/12/15 01:04 [entrez]
PHST- 2020/12/16 06:00 [pubmed]
PHST- 2021/06/22 06:00 [medline]
PHST- 2020/12/09 00:00 [pmc-release]
AID - cells9122647 [pii]
AID - cells-09-02647 [pii]
AID - 10.3390/cells9122647 [doi]
PST - epublish
SO  - Cells. 2020 Dec 9;9(12):2647. doi: 10.3390/cells9122647.