Article stat3 is an upstream regulator of granzyme g in the maternal-to-zygotic transition of mouse embryos
Journal
International Journal of Molecular Sciences
Journal Volume
22
Journal Issue
1
Pages
1-19
Date Issued
2021
Author(s)
Abstract
The maternal-to-zygotic transition (MZT), which controls maternal signaling to synthesize zygotic gene products, promotes the preimplantation development of mouse zygotes to the twocell stage. Our previous study reported that mouse granzyme g (Gzmg), a serine-type protease, is required for the MZT. In this study, we further identified the maternal factors that regulate the Gzmg promoter activity in the zygote to the two-cell stage of mouse embryos. A full-length Gzmg promoter from mouse genomic DNA, FL-pGzmg (?1696~+28 nt), was cloned, and four deletion constructs of this Gzmg promoter, ?1-pGzmg (?1369~+28 nt), ?2-pGzmg (?939~+28 nt), ?3-pGzmg (?711~+28 nt) and ?4-pGzmg (?417~+28 nt), were subsequently generated. Different-sized Gzmg promoters were used to perform promoter assays of mouse zygotes and two-cell stage embryos. The results showed that ?4-pGzmg promoted the highest expression level of the enhanced green fluorescent protein (EGFP) reporter in the zygotes and two-cell embryos. The data suggested that time-specific transcription factors upregulated Gzmg by binding cis-elements in the ?417~+28-nt Gzmg promoter region. According to the results of the promoter assay, the transcription factor binding sites were predicted and analyzed with the JASPAR database, and two transcription factors, signal transducer and activator of transcription 3 (STAT3) and GA-binding protein alpha (GABPα), were identified. Furthermore, STAT3 and GABPα are expressed and located in zygote pronuclei and two-cell nuclei were confirmed by immunofluorescence staining; however, only STAT3 was recruited to the mouse zygote pronuclei and two-cell nuclei injected with the ?4-pGzmg reporter construct. These data indicated that STAT3 is a maternal transcription factor and may upregulate Gzmg to promote the MZT. Furthermore, treatment with a STAT3 inhibitor, S3I-201, caused mouse embryonic arrest at the zygote and two-cell stages. These results suggest that STAT3, a maternal protein, is a critical transcription factor and regulates Gzmg transcription activity in preimplantation mouse embryos. It plays an important role in the maternal-to-zygotic transition during early embryonic development. ? 2021 by the authors. Licensee MDPI, Basel, Switzerland.
Subjects
3 (6 bromo 2 pyridinyl) 2 cyano n (1 phenylethyl)acrylamide; 4 [[(4 methylphenyl)sulfonyloxy]acetamido]salicylic acid; chorionic gonadotropin; enhanced green fluorescent protein; estrogen receptor alpha; estrogen receptor beta; forkhead box f2; GA binding protein; ga binding protein alpha; genomic DNA; granzyme; granzyme g; homeobox protein Meis1; hypoxia inducible factor 1alpha; hypoxia inducible factor 1beta; messenger RNA; myogenic factor; plasmid DNA; STAT3 protein; t cell acute leukemia protein 1; tea domain transcription factor 1; transcription factor; transcription factor FEV; transcription factor GATA 1; transcription factor Sox9; unclassified drug; granzyme; green fluorescent protein; STAT3 protein; Stat3 protein, mouse; transcription factor; animal cell; animal experiment; Article; binding site; cell cycle arrest; cell maturation; controlled study; data base; embryo; embryo development; female; immunofluorescence; maternal to zygotic transition; molecular biology; mouse; nonhuman; phase transition; preimplantation embryo; promoter region; protein expression level; staining; upregulation; zygote; animal; blastocyst; cell nucleus; embryo development; gene expression regulation; genetics; Institute for Cancer Research mouse; male; physiology; pregnancy; transcription initiation; zygote; Animals; Blastocyst; Cell Nucleus; Embryonic Development; Female; Gene Expression Regulation, Developmental; Granzymes; Green Fluorescent Proteins; Male; Mice; Mice, Inbred ICR; Pregnancy; Promoter Regions, Genetic; STAT3 Transcription Factor; Transcription Factors; Transcriptional Activation; Zygote
SDGs
Other Subjects
3 (6 bromo 2 pyridinyl) 2 cyano n (1 phenylethyl)acrylamide; 4 [[(4 methylphenyl)sulfonyloxy]acetamido]salicylic acid; chorionic gonadotropin; enhanced green fluorescent protein; estrogen receptor alpha; estrogen receptor beta; forkhead box f2; GA binding protein; ga binding protein alpha; genomic DNA; granzyme; granzyme g; homeobox protein Meis1; hypoxia inducible factor 1alpha; hypoxia inducible factor 1beta; messenger RNA; myogenic factor; plasmid DNA; STAT3 protein; t cell acute leukemia protein 1; tea domain transcription factor 1; transcription factor; transcription factor FEV; transcription factor GATA 1; transcription factor Sox9; unclassified drug; granzyme; green fluorescent protein; STAT3 protein; Stat3 protein, mouse; transcription factor; animal cell; animal experiment; Article; binding site; cell cycle arrest; cell maturation; controlled study; data base; embryo; embryo development; female; immunofluorescence; maternal to zygotic transition; molecular biology; mouse; nonhuman; phase transition; preimplantation embryo; promoter region; protein expression level; staining; upregulation; zygote; animal; blastocyst; cell nucleus; embryo development; gene expression regulation; genetics; Institute for Cancer Research mouse; male; physiology; pregnancy; transcription initiation; zygote; Animals; Blastocyst; Cell Nucleus; Embryonic Development; Female; Gene Expression Regulation, Developmental; Granzymes; Green Fluorescent Proteins; Male; Mice; Mice, Inbred ICR; Pregnancy; Promoter Regions, Genetic; STAT3 Transcription Factor; Transcription Factors; Transcriptional Activation; Zygote
Type
journal article