1.Mechanisms of alternative splicing in regulating plant flowering: a review.
Huanhuan LU ; Qinlin DENG ; Mengdan WU ; Zhimin WANG ; Dayong WEI ; Hebing WANG ; Huafeng XIANG ; Hongcheng ZHANG ; Qinglin TANG
Chinese Journal of Biotechnology 2021;37(9):2991-3004
Flowering is a critical transitional stage during plant growth and development, and is closely related to seed production and crop yield. The flowering transition is regulated by complex genetic networks, whereas many flowering-related genes generate multiple transcripts through alternative splicing to regulate flowering time. This paper summarizes the molecular mechanisms of alternative splicing in regulating plant flowering from several perspectives, future research directions are also envisioned.
Alternative Splicing/genetics*
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Arabidopsis/metabolism*
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Arabidopsis Proteins/genetics*
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Flowers/genetics*
2.Roles of alternative splicing in infectious diseases: from hosts, pathogens to their interactions.
Mengyuan LYU ; Hongli LAI ; Yili WANG ; Yanbing ZHOU ; Yi CHEN ; Dongsheng WU ; Jie CHEN ; Binwu YING
Chinese Medical Journal 2023;136(7):767-779
Alternative splicing (AS) is an evolutionarily conserved mechanism that removes introns and ligates exons to generate mature messenger RNAs (mRNAs), extremely improving the richness of transcriptome and proteome. Both mammal hosts and pathogens require AS to maintain their life activities, and inherent physiological heterogeneity between mammals and pathogens makes them adopt different ways to perform AS. Mammals and fungi conduct a two-step transesterification reaction by spliceosomes to splice each individual mRNA (named cis -splicing). Parasites also use spliceosomes to splice, but this splicing can occur among different mRNAs (named trans -splicing). Bacteria and viruses directly hijack the host's splicing machinery to accomplish this process. Infection-related changes are reflected in the spliceosome behaviors and the characteristics of various splicing regulators (abundance, modification, distribution, movement speed, and conformation), which further radiate to alterations in the global splicing profiles. Genes with splicing changes are enriched in immune-, growth-, or metabolism-related pathways, highlighting approaches through which hosts crosstalk with pathogens. Based on these infection-specific regulators or AS events, several targeted agents have been developed to fight against pathogens. Here, we summarized recent findings in the field of infection-related splicing, including splicing mechanisms of pathogens and hosts, splicing regulation and aberrant AS events, as well as emerging targeted drugs. We aimed to systemically decode host-pathogen interactions from a perspective of splicing. We further discussed the current strategies of drug development, detection methods, analysis algorithms, and database construction, facilitating the annotation of infection-related splicing and the integration of AS with disease phenotype.
Animals
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Alternative Splicing/genetics*
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RNA Splicing
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Spliceosomes/metabolism*
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RNA, Messenger/metabolism*
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Communicable Diseases/genetics*
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Mammals/metabolism*
3.A progress toward research on alternative splicing of genes in tumor cells.
Chinese Journal of Medical Genetics 2006;23(2):177-180
Alternative splicing of pre-mRNA is an important mechanism for regulating gene function at the post-transcription level and for producing proteomic diversity in higher eukaryotes. The alternative splicing is regulated by the interaction between diverse cis-acting elements and trans-acting factors. Alternative splicing events of oncogenes, tumor suppressor genes and metastasis suppressor genes are associated with the initiation and development of human neoplasms. The protein isoforms sourced from alternative splicing take part in regulating the gene transcription, cell cycle, apoptosis of cells, and playing a role in tumor growth. It is possible for molecular therapy to target directly isoforms of protein produced by alternative splicing or to interfere with the process of alternative splicing.
Alternative Splicing
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genetics
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Humans
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Neoplasms
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genetics
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RNA Precursors
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metabolism
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RNA, Neoplasm
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analysis
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Transcription, Genetic
4.Genetic study of a family of neuronal ceroid lipofuscinosis caused by a heterozygous mutation of gene.
Tie LOU ; Yingzhi HUANG ; Minyue DONG
Journal of Zhejiang University. Medical sciences 2019;48(4):373-377
OBJECTIVE:
To analyze the genetic cause of a family with autosomal recessive neuronal ceroid lipofuscinoses (NCL).
METHODS:
The proband was screened for mutations within the coding region of the candidate genes through high-throughput targeted sequencing. Potential causative mutations were verified by PCR and Sanger sequencing in the proband and his parents. RT-PCR and TA clone sequencing were performed to investigate whether the mRNAs were abnormally spliced.
RESULTS:
The sequencing results revealed compound heterozygous mutations of :c.486+2T>C and c.486+4A>T, which were respectively inherited from his parents. RT-PCR and TA cloning sequencing suggested that the mRNAs were abnormally spliced in two forms due to both mutations.
CONCLUSIONS
The compound heterozygous mutations of :c.486+2T>C and c.486+4A>T are possibly the genetic causes of the NCL family. Detection of the novel mutation has extended mutation spectrum of .
Alternative Splicing
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Female
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Humans
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Male
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Membrane Proteins
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genetics
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Mutation
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Neuronal Ceroid-Lipofuscinoses
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genetics
5.Synonymous variants of the ATP7B gene may cause abnormal splicing of mRNA by affecting the exonic splicing enhancers.
Xiaoying ZHOU ; Bixia ZHENG ; Zhifeng LIU ; Yu JIN
Chinese Journal of Medical Genetics 2020;37(11):1236-1240
OBJECTIVE:
To explore the effect of rare synonymous variants of the ATP7B gene on the splicing of its precursor mRNA.
METHODS:
A total of 248 rare synonymous variants with allelic frequency of <0.005 were retrieved from the ExAc database. Human Splicing Finder (HSF) was used to predict their effect on the splicing of precursor mRNA. And ESE Finder 3.0 was used to predict the effect of such variants on the binding ability of SR protein family. Rare synonymous variants affecting the binding of two or more SR proteins were selected and verified with an in vitro mini gene splicing report system.
RESULTS:
HSF analysis indicated that 136 of the 248 rare synonymous variants may destroy the exonic splicing enhancer (ESE) motif. Analysis using ESE Finder 3.0 indicated that 19 of them may affect the binding of two or more SR proteins at the same time. In vitro mini gene experiment confirmed that the c.1620C>T (p.L540L) and c.3888C>T (p.A1296A) variants could lead to abnormal splicing of the corresponding exons, resulting in complete skipping of exon 4 and 25% increase in the skipping of exon 18, respectively.
CONCLUSION
Synonymous variants may affect the splicing of precursor mRNA in various ways, particularly the destruction of ESE motif. This study confirmed that the c.1620C>T (p.L540L) and c.3888C>T (p.A1296A) variants can affect the mRNA splicing of the ATP7B gene, resulting in skipping of corresponding exons, which may provide a basis for genetic diagnosis and consultation of carriers.
Alternative Splicing
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Copper-Transporting ATPases/genetics*
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Enhancer Elements, Genetic
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Exons
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Gene Frequency
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Humans
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RNA, Messenger/genetics*
6.The research progress of fibronectin EDA's expression and functions.
Journal of Forensic Medicine 2003;19(3):190-inside backcover
7.Alternative Splicing Analysis of LACTB Gene and Expression Characteristics of Different Transcripts in Leukemia Cell Lines.
Ze-Ying LIU ; Fang YANG ; Wei NIE ; Zhi-Qiang YAN ; Qian-Yun SHI ; Bin YUAN ; Li-Rong LIU
Journal of Experimental Hematology 2021;29(4):1019-1027
OBJECTIVE:
To detect the expression of different transcripts of lactamase β(LACTB) gene in leukemic cell lines.
METHODS:
NCBI website and DNAstar software were used to detect the Bioinformatics analysis of LACTB. The expression of different transcripts of LACTB gene in leukemic cell lines (THP-1, HL60, K562, U937, Jurkat and Raji) was detected by reverse transcription PCR (RT-PCR), DNA and clone sequencing; the expression of different transcripts of LACTB gene in leukemic cell lines was detected by Quantitative Real-time PCR.
RESULTS:
There were a variety of splicing isomers in LACTB, and it could produce a variety of protein isomers with conserved N-terminal and different C-terminal, moreover, there were many splice isoforms of LACTB in leukemia cell lines, and there were different expression patterns in different cell lines, including XR1, V1, V2 and V3. The expression of total LACTB showed high in HL60 cells, while low in Raji cells, and the difference was statistically significant (P<0.05). The V1 was high expression in U937 cells but low in Raji cells, and the difference was statistically significant (P<0.05). V2 was high expression in HL60 cells but lowly in Raji cells, and the difference was statistically significant (P<0.05). The expression of V3 was low in THP-1 cells, which was significantly different as compared with that in normal bone marrow (P<0.05).
CONCLUSION
The reaserch found that there are many splice isomers of LACTB in leukemic cell lines, and there are different expression patterns in different cell lines.
Alternative Splicing
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HL-60 Cells
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Humans
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Leukemia/genetics*
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Membrane Proteins/genetics*
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Mitochondrial Proteins/genetics*
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RNA Splicing
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U937 Cells
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beta-Lactamases/genetics*
8.Relationship between the long type PML-RAR alpha and the prognosis of patients with acute promyelocytic leukemia.
Yan-Hong TAN ; Hong-Wei WANG ; Zhi-Ping ZHANG ; Wei LIU ; Hui-Min GUO ; Lei ZHU ; Li ZHANG
Chinese Journal of Hematology 2008;29(1):44-47
OBJECTIVETo explore the relationship between long (L) type PML-RAR alpha fusion gene and the prognosis of patients with acute promyelocytic leukemia (APL).
METHODSPML-RAR alpha fusion gene was detected by RT-PCR in 33 APL patients. The optical density of three bands including E5 (+) E6 (+) 636 bp (no deletion), E5 (-) E6 (+) 492 bp (exon 5 deleted) and E5 (-) E6 (-) 232 bp (both exon 5 and exon 6 deleted) was measured by a UVP analysis system and their relative proportions were calculated. The relative expression level of each splicing band, initial WBC count and age were statistically (for single and multi-factor analysis) analyzed with prognosis.
RESULTSThe relative expressions of E5 (-) E6 (+) and E5 (-) E6 (-) in death group were obviously different from that in first complete remission (CR1) group (P < 0.01), but do not for E5 (+) E6 (+) (P > 0.05). The relative expression levels of E5 (-) E6 (+), E5 (-) E6 (-) and E5 (+) E6 (+) were 0.23 +/- 0.12, 0.58 +/- 0.18, 0.20 +/- 0.09 in death group, and 0.45 +/- 0.16, 0.23 +/- 0.12, 0.31 +/- 0.16 in CR1 group, respectively. Initial WBC count and age was no difference between the two groups (P > 0.05). Logistic regression analysis showed that the expression of E5 (-) E6 (+) had no effect on the prognosis (B = 3.475, P = 0.492), but the expression of E5 (-) E6 (-) showed a negative correlation with prognosis (B = -19.660, P = 0.046).
CONCLUSIONSThe high expression of E5 (-) E6 (-) is correlated with the poor prognosis for patients with APL.
Adolescent ; Adult ; Alternative Splicing ; Female ; Humans ; Leukemia, Promyelocytic, Acute ; genetics ; Male ; Middle Aged ; Oncogene Proteins, Fusion ; genetics ; Prognosis ; Young Adult
9.Expression of alternatively spliced human tissue factor in acute leukemia cells.
Yi FANG ; Jia-Yi CAI ; Ji-Hua ZHONG ; Hua ZHONG ; Hai-Rong WANG ; Fang-Yuan CHEN
Journal of Experimental Hematology 2011;19(2):288-292
The high expression of tissue factor (TF) is related to the coagulation disorder in acute leukemia. TF in blood circulation is mainly expressed in cells, microparticles (MP) and alternatively spliced human tissue factor (asHTF). To elucidate the role of TF in the coagulation disorder of acute myeloid leukemia (AML), RT-PCR was performed on 6 common AML cell lines NB4, HL-60, Kasumi-1, U937, K562 and THP-1. The results showed that only NB4 and U937 cells expressed baseline full-length TF and asHTF which were proved by sequencing. The flow cytometric detection, TF activity and TF antigen tests in NB4 and U937 cells revealed that the asHTF was expressed in trace amount and almost had no activity, while the TF antigen and activity in microparticles were significantly higher than that in asHTF. It is concluded that asHTF may play an unimportant role in the coagulation disorder of AML. Microparticle associated tissue factor (MP-TF) is the predominant source of TF activity released from AML cells.
Alternative Splicing
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HL-60 Cells
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Humans
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Leukemia, Promyelocytic, Acute
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genetics
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metabolism
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Thromboplastin
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genetics
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metabolism
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Tumor Cells, Cultured
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U937 Cells
10.Novel splicing variant of the human orphan nuclear receptor Nurr1 gene.
Chinese Medical Journal 2004;117(6):899-902
BACKGROUNDNurr1 is a member of the nuclear receptor superfamily of transcription factors. The objective of the present study was to identify novel splicing variants of the gene in neuronal and non-neuronal tissues and determine their functions.
METHODSReverse transcription-polymerase chain reaction (RT-PCR) analysis was used to screen for Nurr1 splice variants in the adult human central nervous system (CNS) and in other tissues such as lymphocytes, and liver, muscle, and kidney cells. Functional assays of the variants were performed by measuring Nurr1 response element (NuRE) transcriptional activity in vitro.
RESULTSIn this study, the authors identified a novel splicing variant of Nurr1 within exon 5, found in multiple adult human tissues, including lymphocytes, and liver, muscle, and kidney cells, but not in the brain or spinal cord. Sequencing analysis showed the variant has a 75 bp deletion between nucleotides 1402 and 1476. A functional assay of the Nurr1-c splicing variant, performed by measuring NuRE transcriptional activity in vitro, detected a 39% lower level of luciferase (LUC) activity (P < 0.05).
CONCLUSIONA novel splicing variant of Nurr1 exists in human non-neuronal tissues and functional assays suggest that the variant may act as an alternate transcription regulator.
Adult ; Alternative Splicing ; DNA-Binding Proteins ; analysis ; genetics ; Humans ; Nuclear Receptor Subfamily 4, Group A, Member 2 ; Transcription Factors ; analysis ; genetics