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MACF1 Promotes Osteoblast Differentiation by Sequestering Wnt Signaling Repressors in Cytoplasm [RNA-seq]

ABSTRACT: MACF1 Promotes Osteoblast Differentiation by Sequestering Wnt Signaling Repressors in Cytoplasm [RNA-seq]

PROVIDER: PRJNA492300 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR7883050_1.fastq.gz	Fastqsanger.gz
	SRR7883050_2.fastq.gz	Fastqsanger.gz
	SRR7883060_1.fastq.gz	Fastqsanger.gz
	SRR7883060_2.fastq.gz	Fastqsanger.gz
	SRR7883051_1.fastq.gz	Fastqsanger.gz

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MACF1 promotes osteoblast differentiation by sequestering repressors in cytoplasm

Project description:MACF1 promotes osteoblast differentiation by sequestering repressors in cytoplasm

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MACF1 Promotes Osteoblast Differentiation by Sequestering Wnt Signaling Repressors in Cytoplasm [ChIP-seq]

Project description:Osteoblast differentiation leading to bone formation requires a coordinated transcriptional program. We have recently demonstrated that microtubule actin crosslinking factor 1 (MACF1) promotes osteoblast differentiation, suggesting a key role in regulating early-phase osteoblast differentiation. Here, we showed that the early-phase osteoblast differentiation transcriptome dynamics was regulated by MACF1 and the transcription of TCF7/LEF1, key effectors of Wnt signaling that is important for osteoblast differentiation was suppressed by MACF1 knockdown. Co-IP and Protein mass spectrometry revealed that MACF1 interacted with a known and two previously unknown repressors of TCF7/LEF1, DKK1, CDK12 and MEAF6. ChIP-seq analysis of MACF1-associated promoters further revealed that MACF1 interacted with transcription factors TCF12 and E2F6, which also suppressed the transcription of TCF7/LEF1. Furthermore, all these four MACF-interacted proteins inhibited osteoblast differentiation. By studying the underlying mechanism, we found that cytoplasmic-nuclear localization of MACF1 was dependent on its level and the cytoplasmic-nuclear localization of TCF12 and E2F6 was regulated by MACF1 localization. In addition, MACF1 oppositely regulated the transcription activity of TCF12 and TCF7. Current study, for the first time to our knowledge, suggest that MACF1 acts as a sponge of osteoblast differentiation repressors to promote osteoblast differentiation, and indicate a novel mechanism for regulating the cellular location of transcription factors by a protein associated with microtubule and actin.

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MACF1 Promotes Osteoblast Differentiation by Sequestering Wnt Signaling Repressors in Cytoplasm [RNA-seq]

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MACF1 Promotes Osteoblast Differentiation by Sequestering Wnt Signaling Repressors in Cytoplasm [ChIP-seq]

Project description:MACF1 Promotes Osteoblast Differentiation by Sequestering Wnt Signaling Repressors in Cytoplasm [ChIP-seq]

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MACF1 promotes osteoblast differentiation by sequestering repressors in cytoplasm.

Project description:Osteoblast differentiation leading to bone formation requires a coordinated transcriptional program. Osteoblastic cells with low level of microtubule actin crosslinking factor 1 (MACF1) show reduced osteoblast differentiation ability, however, the comprehensive mechanism of MACF1's action remains unexplored. In the current study, we found that MACF1 knockdown suppressed osteoblast differentiation by altering the transcriptome dynamics. We further identified two MACF1-interacted proteins, cyclin-dependent kinase 12 (CDK12) and MYST/Esa1-associated factor 6 (MEAF6), and two MACF1-interacted transcription factors (TFs), transcription factor 12 (TCF12) and E2F transcription factor 6 (E2F6), which repress osteoblast differentiation by altering the expression of osteogenic TFs and genes. Moreover, we found that MACF1 regulated cytoplasmic-nuclear localization of itself, TCF12 and E2F6 in a concentration-dependent manner. MACF1 oppositely regulates the expression of TCF12 and transcription factor 7 (TCF7), two TFs that drive osteoblast differentiation to opposite directions. This study reveals that MACF1, a cytoskeletal protein, acts as a sponge for repressors of osteoblast differentiation to promote osteoblast differentiation and contributes to a novel mechanistic insight of osteoblast differentiation and transcription dynamics.

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MACF1 promotes osteoblast differentiation by sequestering repressors in cytoplasm

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Project description:MC3T3-E1 cells were transfected with short hairpin RNA (shRNA) specifically targeting the murine MACF1 lentivirus vector or with scrambled shRNA, and the stably transfected cell lines were selected using puromycin. After 15 days of selection, all cells were collected for further study.

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