Runx1 exon 6–related alternative splicing isoforms differentially regulate hematopoiesis in mice

Komeno, Yukiko; Yan, Ming; Matsuura, Shinobu; Lam, Kentson; Lo, Miao Chia; Huang, Yi-Jou; Tenen, Daniel G.; Downing, James R.; Zhang, Dong Er

doi:10.1182/blood-2013-08-521252

Cited by 34 publications

(54 citation statements)

References 50 publications

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“…The virus infection procedure was performed as described previously (27). For general retrovirus infection, 293T cells were transfected with 5 g of retrovirus vectors and with 5 g of Ecopac packaging vector using polyethylenimine (Polysciences Inc.).…”

Section: Methodsmentioning

confidence: 99%

“…293T cells, mouse fetal liver (FL) cells (embryonic day 12.5 [E12.5] to E14.5), total bone marrow (BM) cells, and lineage-depleted BM cells (sorted by using a Lineage depletion kit from Miltenyi) were cultured as described before (27). To make a single-cell suspension of FL cells, FLs were sheared in PBS by pipetting, passed through 40-mpore-size cell strainers, and treated with ammonium-chloride-potassium (ACK) buffer (150 mM NH 4 Cl, 1 mM KHCO 3 , 0.1 mM EDTA) when necessary.…”

Section: Methodsmentioning

confidence: 99%

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SRSF2 Is Essential for Hematopoiesis, and Its Myelodysplastic Syndrome-Related Mutations Dysregulate Alternative Pre-mRNA Splicing

Komeno

Huang

Qiu

et al. 2015

Molecular and Cellular Biology

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dMyelodysplastic syndromes (MDS) are a group of neoplasms characterized by ineffective myeloid hematopoiesis and various risks for leukemia. SRSF2, a member of the serine/arginine-rich (SR) family of splicing factors, is one of the mutation targets associated with poor survival in patients suffering from myelodysplastic syndromes. Here we report the biological function of SRSF2 in hematopoiesis by using conditional knockout mouse models. Ablation of SRSF2 in the hematopoietic lineage caused embryonic lethality, and Srsf2-deficient fetal liver cells showed significantly enhanced apoptosis and decreased levels of hematopoietic stem/progenitor cells. Induced ablation of SRSF2 in adult Mx1-Cre Srsf2 flox/flox mice upon poly(I):poly(C) injection demonstrated a significant decrease in lineage ؊ Sca ؉ c-Kit ؉ cells in bone marrow. To reveal the functional impact of myelodysplastic syndromes-associated mutations in SRSF2, we analyzed splicing responses on the MSD-L cell line and found that the missense mutation of proline 95 to histidine (P95H) and a P95-to-R102 in-frame 8-amino-acid deletion caused significant changes in alternative splicing. The affected genes were enriched in cancer development and apoptosis. These findings suggest that intact SRSF2 is essential for the functional integrity of the hematopoietic system and that its mutations likely contribute to development of myelodysplastic syndromes. Multiple classes of genetic aberrations have been suggested as the cause of myelodysplastic syndromes (MDS) (1, 2), including mutations in signal transduction, transcription factors, and epigenetic modifiers (3-5). Interestingly, recent genomewide sequencing studies revealed that mutations in genes encoding splicing factors are commonly associated with MDS and other hematological malignancies (6-15). One of these newly identified genes codes for the SRSF2 splicing factor (also known as SC35), and its mutations have been linked to poor survival among MDS patients (16,17). Most of the SRSF2 mutations occurred at proline 95, and the majority of these mutations changed this proline to histidine (P95H); less-frequent changes to leucine (P95L) and arginine (P95R) and in-frame deletion of 8 amino acids (aa) from P95 to R102 (⌬8aa) have also been reported previously (6,16,(18)(19)(20). However, the causal effect of these mutations on MDS development remains to be established. SRSF2 is one of the founding members of the serine/argininerich (SR) protein family of splicing factors (21). It is involved in both constitutive and regulated splicing. Homozygous germ line Srsf2 knockout (KO) mice are embryonically lethal (22), and conditional knockout (cKO) mice display various tissue-specific phenotypes (22-24). Importantly, Srsf2 downregulation in mouse embryonic fibroblasts results in G 2 /M cell cycle arrest and genomic instability (23). To date, systematic analysis of SRSF2 function in the blood system has not been reported except for its requirement in T cell development (24). Given the tight link of Srsf2 mutations to MDS, we ai...

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

SRSF2 Is Essential for Hematopoiesis, and Its Myelodysplastic Syndrome-Related Mutations Dysregulate Alternative Pre-mRNA Splicing

Komeno

Huang

Qiu

et al. 2015

Molecular and Cellular Biology

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“…A similar short isoform lacking the transactivation domain was described in mice, albeit with a different 39 exon (Figure 3). 127,128 Interestingly, the mouse short Runx1 isoform had a biological role similar to that of the human RUNX1a in that it positively regulated the size of the HSC pool. 127 The full-length P1-and P2-derived Runx1c and Runx1b protein isoforms are identical apart from their N terminus.…”

Section: Structure Of Mammalian Runx Genes and Alternatively Spliced mentioning

confidence: 99%

“…127,128 Interestingly, the mouse short Runx1 isoform had a biological role similar to that of the human RUNX1a in that it positively regulated the size of the HSC pool. 127 The full-length P1-and P2-derived Runx1c and Runx1b protein isoforms are identical apart from their N terminus. In human and mouse, there are 32 and 19 unique RUNX1c N-terminal amino acids, respectively, and 5 unique RUNX1b ones.…”

Section: Structure Of Mammalian Runx Genes and Alternatively Spliced mentioning

confidence: 99%

Runx transcription factors in the development and function of the definitive hematopoietic system

Bruijn

Dzierzak

2017

Blood

153

103

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The Runx family of transcription factors (Runx1, Runx2, and Runx3) are highly conserved and encode proteins involved in a variety of cell lineages, including blood and blood-related cell lineages, during developmental and adult stages of life. They perform activation and repressive functions in the regulation of gene expression. The requirement for Runx1 in the normal hematopoietic development and its dysregulation through chromosomal translocations and loss-of-function mutations as found in acute myeloid leukemias highlight the importance of this transcription factor in the healthy blood system. Whereas another review will focus on the role of Runx factors in leukemias, this review will provide an overview of the normal regulation and function of Runx factors in hematopoiesis and focus particularly on the biological effects of Runx1 in the generation of hematopoietic stem cells. We will present the current knowledge of the structure and regulatory features directing lineage-specific expression of Runx genes, the models of embryonic and adult hematopoietic development that provide information on their function, and some of the mechanisms by which they affect hematopoietic function.

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“…6,7 Of 3 major isotypes, we focused on both functional full-length isoforms RUNX1b/c (lacking exon 7a and including exon 7b and 8), and the short isoform, RUNX1a (including exon 7a, which has a stop codon), which is found only in primates. 8 RUNX1b and RUNX1c are functionally identical and RUNX1b is broadly expressed, 9 whereas RUNX1a has a dominant-negative effect on RUNX1b. 10 RUNX1a overexpression has been reported in AML.…”

Section: Introductionmentioning

confidence: 99%

Overexpression of RUNX1 short isoform has an important role in the development of myelodysplastic/myeloproliferative neoplasms

et al. 2017

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Runx1 exon 6–related alternative splicing isoforms differentially regulate hematopoiesis in mice

Cited by 34 publications

References 50 publications

SRSF2 Is Essential for Hematopoiesis, and Its Myelodysplastic Syndrome-Related Mutations Dysregulate Alternative Pre-mRNA Splicing

SRSF2 Is Essential for Hematopoiesis, and Its Myelodysplastic Syndrome-Related Mutations Dysregulate Alternative Pre-mRNA Splicing

Runx transcription factors in the development and function of the definitive hematopoietic system

Overexpression of RUNX1 short isoform has an important role in the development of myelodysplastic/myeloproliferative neoplasms

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