PC7 and the related proteases Furin and Pace4 regulate E-cadherin function during blastocyst formation

Bessonnard, Sylvain; Mesnard, Daniel; Constam, Daniel

doi:10.1083/jcb.201503042

Cited by 20 publications

(31 citation statements)

References 68 publications

(107 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…S8B; ref. 30) and E-cadherin, which has been shown to be a furin and PC7 substrate (31). For both IGF1R and ITGA6, only furin knockdown and CMK treatments prevented the processing of their pro-forms, whereas PACE4 knockdown and the C23 PACE4 inhibitor had no effect, further supporting its selectivity toward PACE4 even in cells with concentrations above the selectivity range (K i : low nanomolar).…”

Section: Identification Of Gdf-15 As a Specific Pace4 Substrate And Pmentioning

confidence: 79%

PACE4 Undergoes an Oncogenic Alternative Splicing Switch in Cancer

et al. 2017

View full text Add to dashboard Cite

Inhibition of PACE4, a proprotein convertase that is overexpressed in prostate cancer, has been shown to block cancer progression in an androgen-independent manner. However, the basis for its overexpression and its growth-inhibitory effects are mitigated and uncertain. Here, we report that PACE4 pre-mRNA undergoes DNA methylation-sensitive alternative splicing of its terminal exon 3 0 untranslated region, generating an oncogenic, C-terminally modified isoform (PACE4-altCT). We found this isoform to be strongly expressed in prostate cancer cells, where it displayed an enhanced autoactivating process and a distinct intracellular routing that prevented its extracellular secretion. Together, these events led to a dramatic increase in processing of the progrowth differentiation factor pro-GDF15 as the first PACE4 substrate to be identified in prostate cancer. We detected robust expression of PACE4-altCT in other cancer types, suggesting that an oncogenic switch for this proenzyme may offer a therapeutic target not only in advanced prostate cancer but perhaps also more broadly in human cancer. Cancer Res; 77(24); 6863-79. Ó2017 AACR.

show abstract

Section: Identification Of Gdf-15 As a Specific Pace4 Substrate And Pmentioning

confidence: 79%

PACE4 Undergoes an Oncogenic Alternative Splicing Switch in Cancer

et al. 2017

View full text Add to dashboard Cite

show abstract

“…Consistent with the aforementioned studies, we also discovered that cell adhesion-related genes, including CDH1, were up-regulated by KDM6A overexpression in bovine embryos. Importantly, during preimplantation embryo development, adherens and tight junctions coordinately establish the first internal cavity of the embryo to transform the morula into a blastocyst (54); perturbation of these junctions leads to defects in blastocyst formation (55)(56)(57). These results provide a basis for investigating the mechanism of the first cell fate determination during embryonic development in mammals.…”

Section: Discussionmentioning

confidence: 96%

H3K27me3 is an epigenetic barrier while KDM6A overexpression improves nuclear reprogramming efficiency

et al. 2019

View full text Add to dashboard Cite

Aberrant epigenetic reprogramming is a major factor of developmental failure of cloned embryos. Histone H3 lysine 27 trimethylation (H3K27me3), a histone mark for transcriptional repression, plays important roles in mammalian embryonic development and induced pluripotent stem cell (iPSC) generation. The global loss of H3K27me3 marks may facilitate iPSC generation in mice and humans. However, the H3K27me3 level and its role in bovine somatic cell nuclear transfer (SCNT) reprogramming remain poorly understood. Here, we show that SCNT embryos exhibit global H3K27me3 hypermethylation from the 2‐ to 8‐cell stage and that its removal by ectopically expressed H3K27me3 lysine demethylase (KDM)6A greatly improves nuclear reprogramming efficiency. In contrast, H3K27me3 reduction by H3K27me3 methylase enhancer of zeste 2 polycomb repressive complex knockdown or donor cell treatment with the enhancer of zeste 2 polycomb repressive complex—selective inhibitor GSK343 suppressed blastocyst formation by SCNT embryos. KDM6A overexpression enhanced the transcription of genes involved in cell adhesion and cellular metabolism and X‐linked genes. Furthermore, we identified methyl‐CpG‐binding domain protein 3‐like 2, which was reactivated by KDM6A, as a factor that is required for effective reprogramming in bovines. These results show that H3K27me3 functions as an epigenetic barrier and that KDM6A overexpression improves SCNT efficiency by facilitating transcriptional reprogramming.—Zhou, C., Wang, Y., Zhang, J., Su, J., An, Q., Liu, X., Zhang, M., Wang, Y., Liu, J., Zhang, Y. H3K27me3 is an epigenetic barrier while KDM6A overexpression improves nuclear reprogramming efficiency. FASEB J. 33, 4638–4652 (2019). http://www.fasebj.org

show abstract

“…HEK293T cells used in our screening assays express PC2, furin, PACE4, PC5/6 and PC7 , which at their endogenous levels fail to cleave BPCS beyond background levels of u‐BPCS. Next, we examined possible processing of BPCS after overexpression of the nonbasic PC SKI‐1/S1P that recognizes cleavage sites Basic‐X‐(hydrophobic)‐X↓ .…”

Section: Resultsmentioning

confidence: 99%

“…HEK293T cells used in our screening assays express PC2, furin, PACE4, PC5/6 and PC7 [27], which at their endogenous levels fail to cleave BPCS beyond The N-terminal GLuc is linked to the transmembrane and cytosolic domains of sortilin-1 via the POMC-derived sequence NSSSSGSSGAGQKR↓EDVSAGEDCGPLPEGGP, followed by a V5 peptide tag for detection. The scissile bond behind KR is indicated by scissors and the putative N-glycosylation motif (NSS) underlined.…”

Section: Introductionmentioning

confidence: 99%

A novel cell‐based sensor detecting the activity of individual basic proprotein convertases

et al. 2019

Self Cite

View full text Add to dashboard Cite

The basic proprotein convertases (PCs) furin, PC1/3, PC2, PC5/6, PACE4, PC4, and PC7 are promising drug targets for human diseases. However, developing selective inhibitors remains challenging due to overlapping substrate recognition motifs and limited structural information. Classical drug screening approaches for basic PC inhibitors involve homogeneous biochemical assays using soluble recombinant enzymes combined with fluorogenic substrate peptides that may not accurately recapitulate the complex cellular context of the basic PC–substrate interaction. Herein we report basic PC sensor (BPCS), a novel cell‐based molecular sensor that allows rapid screening of candidate inhibitors and their selectivity toward individual basic PCs within mammalian cells. BPCS consists of Gaussia luciferase linked to a sortilin‐1 membrane anchor via a cleavage motif that allows efficient release of luciferase specifically if individual basic PCs are provided in the same membrane. Screening of selected candidate peptidomimetic inhibitors revealed that BPCS can readily distinguish between general and selective PC inhibitors in a high‐throughput screening format. The robust and cost‐effective assay format of BPCS makes it suitable to identify novel specific small‐molecule inhibitors against basic PCs for therapeutic application. Its cell‐based nature will allow screening for drug targets in addition to the catalytically active mature enzyme, including maturation, transport, and cellular factors that modulate the enzyme's activity. This broadened ‘target range’ will enhance the likelihood to identify novel small‐molecule compounds that inhibit basic PCs in a direct or indirect manner and represents a conceptual advantage.

show abstract

PC7 and the related proteases Furin and Pace4 regulate E-cadherin function during blastocyst formation

Abstract: Targeted deletion of PC7 and the related proprotein convertases Furin and Pace4, combined with live imaging of their activities, unmasks their overlapping and complementary functions in morula compaction and ICM formation in mouse blastocysts and in E-cadherin precursor processing.

Cited by 20 publications

References 68 publications

PACE4 Undergoes an Oncogenic Alternative Splicing Switch in Cancer

PACE4 Undergoes an Oncogenic Alternative Splicing Switch in Cancer

H3K27me3 is an epigenetic barrier while KDM6A overexpression improves nuclear reprogramming efficiency

A novel cell‐based sensor detecting the activity of individual basic proprotein convertases

Contact Info

Product

Resources

About