Alternative titles; symbols
HGNC Approved Gene Symbol: ERF
Cytogenetic location: 19q13.2 Genomic coordinates (GRCh38) : 19:42,247,569-42,255,128 (from NCBI)
| Location | Phenotype |
Phenotype MIM number |
Inheritance |
Phenotype mapping key |
|---|---|---|---|---|
| 19q13.2 | Chitayat syndrome | 617180 | Autosomal dominant | 3 |
| Craniosynostosis 4 | 600775 | Autosomal dominant | 3 |
Members of the ETS family of transcription factors, such as ERF, regulate cell proliferation and differentiation. They share a highly conserved DNA-binding domain, the ETS domain, that recognizes the sequence GGAA/T (de Castro et al., 1997). For further information on ETS transcription factors, see ETS1 (164720).
By screening a myelogenous leukemia cell line cDNA library with an ETS-binding site in the ETS2 (164740) promoter, followed by screening a placenta cDNA library, Sgouras et al. (1995) obtained full-length ERF. The deduced 548-amino acid protein contains an N-terminal ETS domain structurally similar to the ETS domains of ELK1 (311040) and SAP1 (ELK4; 600246). It also has a central proline-rich region, 2 putative SH3 interaction sites, and several putative phosphorylation sites. Northern blot analysis detected a 2.7-kb transcript in all tissues and cell lines examined. Western blot analysis showed multiple phosphorylated forms of ERF ranging from 75 to 85 kD.
By RT-PCR of human leukemia cell lines using degenerate oligonucleotides based on the ETS domains of FLI1 (193067) and ERG (165080), followed by screening leukemia cell line and testis cDNA libraries, de Castro et al. (1997) cloned ERF, which they called PE2. The ETS domain of ERF is most similar to the ETS domains of PE1 (ETV3; 164873), FLI1, and ERG. Northern blot analysis detected ERF expression in all human tissues and cell lines examined, with highest levels in heart, testis, ovary, and pancreas.
Liu et al. (1997) cloned mouse Erf. The human and mouse ERF proteins share 98% amino acid identity, with most of the differences in the C-terminal transcriptional repressor domain.
By whole-mount RNA in situ hybridization and RT-PCR, Twigg et al. (2013) observed Erf expression in wildtype mice along the osteogenic margins of developing calvarial bones, in a similar distribution to that seen for the master osteogenic regulator Runx2 (600211).
De Castro et al. (1997) determined that the ERF gene contains 4 exons and spans more than 9 kb. The 5-prime flanking region is GC rich. It has no canonical TATA-binding motif, but contains several binding motifs for AP-1 (see 165160) and CREB (see 123810) factors and a single MYB (189990)-binding site.
Liu et al. (1997) identified a conserved TATA element in mouse and human ERF. Several transcription factor-binding sites are also conserved, including 2 SP1 (189906) sites and an ETS-binding site.
By somatic cell hybrid analysis and FISH, de Castro et al. (1997) mapped the ERF gene to chromosome 19q13.2-q13.31.
Using FISH, Liu et al. (1997) mapped the ERF gene to chromosome 19q13.1. They mapped the mouse Erf gene to a region of chromosome 7 that shares homology of synteny with human chromosome 19q13.
Using transfection assays of HeLa cells, Sgouras et al. (1995) found that ERF repressed the ETS2 promoter more than 30-fold. ERF also repressed the GATA1 (605371) promoter in the presence of its transactivator, GATA1. ERF suppressed the ETS-dependent transforming activity of the gag-myb (189990)-ets fusion oncogene of the avian E26 virus. The level of transcriptional repression by ERF was proportional to its affinity for the ERF target sequence in the promoter. The isolated DNA-binding domain of ERF strongly inhibited the ETS2 promoter, whereas removal of the DNA-binding domain of ERF abrogated its repressor activity. Sgouras et al. (1995) mapped the ERF repressor domain between amino acids 472 and 530. Although the ERF protein level remained constant throughout the cell cycle, ERF phosphorylation status was altered as a function of the cell cycle and after mitogenic stimulation. ERF was hyperphosphorylated in cells transformed by the activated Ha-ras (HRAS; 190020) and v-src (SRC; 190090) genes, and the transcription repressor activity of ERF was decreased after cotransfection with activated Ha-ras or the kinase domain of RAF1 (164760), indicating that ERF activity is probably regulated by the Ras/MAPK pathway. Consistent with the in vivo phosphorylation and inactivation by Ha-ras, ERF was efficiently phosphorylated in vitro by Erk2 (MAPK1; 176948) and CDC2 (116940)/cyclin B (see CCNB1; 123836) kinases at sites similar to those detected in vivo. Mutation of thr526 to ala eliminated a major ERF phosphoprotein in vivo, after phorbol ester induction, and in vitro, after ERK2 phosphorylation. Substitution of thr526 for glu also decreased the repression ability of ERF. Sgouras et al. (1995) concluded that ERF is involved in transcriptional regulation of genes activated during entry into G1 phase.
Using in situ hybridization, Janesick et al. (2013) showed that retinoic acid receptor (RAR; see 180240) signaling induced neuronal differentiation and inhibited proproliferative neural markers in Xenopus embryos. Further analysis identified Erf, Etv3 (164873), and Etv3l (621105) as RAR-responsive repressors expressed in neuroectoderm. Erf or Etv3/Etv3l knockdown inhibited the neural differentiation pathway and expanded expression of neural progenitor markers in Xenopus embryos. Moreover, knockdown of Erf or Etv3l promoted proliferation in the neural plate in Xenopus embryos, whereas Erf overexpression led to an increase of neurons.
Bose et al. (2017) showed that ERF mutations in prostate cancer cause decreased protein stability and mostly occur in tumors without ERG (165080) upregulation. ERF loss recapitulated the morphologic and phenotypic features of ERG gain in normal mouse prostate cells, including expansion of the androgen receptor (AR; 313700) transcriptional repertoire, and ERF had tumor suppressor activity in the same genetic background of PTEN (601728) loss that yields oncogenic activity by ERG. In the more common scenario of ERG upregulation, chromatin immunoprecipitation followed by sequencing indicated that ERG inhibits the ability of ERF to bind DNA at consensus ETS sites both in normal and in cancerous prostate cells. Consistent with a competition model, ERF overexpression blocked ERG-dependent tumor growth, and ERF loss rescued TMPRSS2 (602060)-ERG-positive prostate cancer cells from ERG dependency. Bose et al. (2017) concluded that their data provided evidence that the oncogenicity of ERG is mediated, in part, by competition with ERF, and raised the larger question of whether other gain-of-function oncogenic transcription factors might also inactivate endogenous tumor suppressors.
Craniosynostosis 4
In 12 families with craniosynostosis (CRS4; 600775), Twigg et al. (2013) identified heterozygosity for mutations in the ERF gene (see, e.g., 611888.0001-611888.0005).
Chaudhry et al. (2015) analyzed the ERF gene in 40 patients with genetically undiagnosed multiple suture or sagittal synostosis, and identified 2 heterozygous mutations, one in the start codon (611888.0006) and the other in a splice site (611888.0007), in 2 affected boys. Chaudhry et al. (2015) noted that 2 (5%) of 40 patients in their study population had a pathogenic variant in the ERF gene, similar to the 3% frequency that was observed in a larger cohort (Twigg et al., 2013).
In a cohort of 182 Spanish craniosynostosis probands, Paumard-Hernandez et al. (2015) screened 7 craniosynostosis-associated genes, including the ERF gene, but did not detect any mutations in ERF. The authors suggested that the frequency of ERF mutations might be lower than previously reported.
Chitayat Syndrome
In 5 patients from 4 families with Chitayat syndrome (CHYTS; 617180), Balasubramanian et al. (2017) identified heterozygosity for a recurrent missense mutation in the ERF gene (Y89C; 611888.0008). The authors stated that it was unclear why the Y89C variant produced a different phenotype from that associated with previously reported mutations in the same region of ERF, such as R86C (611888.0003) and R65Q (611888.0004).
Papadaki et al. (2007) found that mouse Erf was expressed throughout embryonic development and adulthood. However, in situ hybridization of developing placenta showed that, after 7.5 days postcoitum, expression of Erf was restricted to extraembryonic ectoderm, and after 9.5 days postcoitum, it was restricted to a subpopulation of labyrinth cells. Erf +/- mice appeared normal and were fertile, but Erf -/- embryos died in utero at day 10 due to severe placenta defects. Erf -/- embryos failed to undergo chorioallantoic attachment and labyrinth development and instead had an expanded chorion layer that failed to further differentiate. These mice also failed to close the ectoplacental cone cavity. Erf -/- placentas had abnormalities in the giant cell and spongiotrophoblast layers. Erf -/- trophoblast stem cells showed delayed differentiation compared with wildtype cells and failed to express specific differentiation markers.
Twigg et al. (2013) generated mice with a conditional Erf allele and observed that mice heterozygous or homozygous for the conditional allele were grossly normal, whereas mice that were compound heterozygous for the conditional allele and a null allele had domed heads that became apparent during the first 3 to 6 weeks of life. Micro-CT scanning showed craniosynostosis affecting multiple calvarial sutures. No other specific skeletal abnormalities were evident. Quantification of transcripts in embryonic day 16.5 calvaria showed up to 2-fold downregulation of multiple osteogenic markers in conditional/null mutants compared to wildtype littermates. Despite this mildly delayed embryonic ossification, by postnatal day 14 there was variable fusion of the cranial sutures of mutant, but not of wildtype, pups.
In 2 brothers with craniosynostosis (CRS4; 600775), Twigg et al. (2013) identified heterozygosity for a c.547C-T transition in exon 4 of the ERF gene, resulting in an arg183-to-ter (R183X) substitution. The mutation was also present in their mother, who had exorbitism and midface hypoplasia suggestive of Crouzon syndrome (123500) but was negative for mutation in the FGFR2 gene (176943); she had inherited the R183X ERF mutation from her mother, who exhibited only macrocephaly. Immunoblotting of fibroblasts or lymphoblastoid cells from affected individuals showed lower expression of full-length ERF compared to wildtype cells. The older brother had craniosynostosis affecting all sutures of the cranial vault, whereas the younger brother had metopic, sagittal, and left coronal synostosis.
In 4 affected individuals from 3 unrelated families with craniosynostosis (CRS4; 600775), Twigg et al. (2013) identified heterozygosity for a 2-bp deletion (c.891_892delAG) in exon 4 of the ERF gene, causing a frameshift predicted to result in a premature termination codon (Gly299ArgfsTer9) within the ERK interaction domain. Haplotype analysis in 2 of the families revealed different microsatellite genotypes. One 10-year-old male proband was diagnosed with syndromic pansynostosis and had a copper-beaten appearance of the skull, hypertelorism, exorbitism, downslanting palpebral fissures, and global developmental delay involving poor attention span and receptive language, and behavioral difficulties. The 9.4-year-old male proband from a second family, who was diagnosed with nonsyndromic craniosynostosis involving the sagittal and bilateral lambdoid sutures, had elevated intracranial pressure preoperatively as well as specific language impairment. In the third family, there were 2 affected sisters diagnosed with syndromic craniosynostosis: the elder, who had features suggestive of Crouzon syndrome (123500) but was negative for mutation in the FGFR2 gene (176943), exhibited bilateral coronal and lambdoid synostosis, hypertelorism, exorbitism, and midface hypoplasia, whereas the younger had bilateral lambdoid involvement with hypertelorism, hyperactivity, and poor concentration. Their mother, who also carried the 2-bp deletion in the ERF gene, exhibited exorbitism and short digits.
In 2 unrelated boys with craniosynostosis (CRS4; 600775), Twigg et al. (2013) identified heterozygosity for a c.256C-T transition in exon 2 of the ERF gene, resulting in an arg86-to-cys (R86C) substitution at a highly conserved residue in the ETS domain. One of the boys, who was diagnosed with nonsyndromic synostosis of the right lambdoid suture, had a Chiari type I malformation (see 118420), macrocephaly, speech and language delay, and middle ear effusions; he had inherited the mutation from his unaffected mother. The other boy, who had been diagnosed with Crouzon (123500)-like syndromic craniosynostosis but was negative for mutation in the FGFR2 gene (176943), had synostosis of the sagittal and bilateral lambdoid sutures with elevated intracranial pressure at the time of his first operative procedure, and exhibited hypertelorism, exorbitism, class I dental malocclusion, broad thumbs and halluces, and a speech articulation disorder with verbal dyspraxia and general motor development dyspraxia. The R86C mutation arose de novo in both the second proband and in the first proband's mother.
In a 9.9-year-old boy with craniosynostosis (CRS4; 600775), Twigg et al. (2013) identified heterozygosity for a c.194G-A transition in exon 2 of the ERF gene, resulting in an arg65-to-gln (R65Q) substitution at a conserved residue in the ETS domain. The boy, who was diagnosed with nonsyndromic craniosynostosis involving the sagittal and right coronal sutures, also had a platelet disorder. He inherited the mutation from his mother, who was macrocephalic without known craniosynostosis.
In a 10-year-old girl with craniosynostosis (CRS4; 600775), Twigg et al. (2013) identified heterozygosity for a c.1270C-T transition in exon 4 of the ERF gene, resulting in a gln424-to-ter (Q424X) substitution. Immunoblotting of fibroblasts or lymphoblastoid cells from affected individuals showed lower expression of full-length ERF compared to wildtype cells. The girl, who had been diagnosed with a Crouzon (123500)-like syndromic craniosynostosis but was negative for mutation in the FGFR2 gene (176943), had synostosis of all sutures of the cranial vault, a Chiari type I malformation (see 118420), copper-beaten appearance of skull, hypertelorism, exorbitism, thoracic and lumbar scoliosis, and broad toes. Her 8-year-old brother, who also carried the mutation, did not have craniosynostosis but did have a Chiari I malformation, as well as hypertelorism, exorbitism, and broad toes. The sibs inherited the Q424X mutation from their mother, who exhibited only hypertelorism, and also had an unspecified cardiac septal defect and von Willebrand disease (see 193400).
In a boy (A1) with craniosynostosis involving all sutures (CRS4; 600775), Chaudhry et al. (2015) identified heterozygosity for a c.1A-G transition in the ERF gene, predicted to silence the start codon and impair proper transcription of ERF. Parental DNA was unavailable for study.
In a boy with bilateral coronal and metopic craniosynostosis (CRS4; 600775), Chaudhry et al. (2015) identified heterozygosity for a c.23-2A-G transition within a conserved splice acceptor site of exon 2, predicted to cause skipping of exon 2 during transcript maturation. Parental DNA was unavailable for study.
In 5 patients from 4 families with Chitayat syndrome (CHYTS; 617180), including the patient originally described by Chitayat et al. (1993), Balasubramanian et al. (2017) identified heterozygosity for a c.266A-G (c.266A-G, NM_006494.2) transition in the ERF gene, resulting in a tyr89-to-cys (Y89C) substitution at a highly conserved residue within the DNA-binding ETS domain. The mutation, which was detected in an affected father and son in 1 of the families, was confirmed to have occurred de novo in the remaining 3 patients. Investigations including CT scans of the skull detected no evidence of craniosynostosis in any of the patients.
Balasubramanian, M., Lord, H., Levesque, S., Guturu, H., Thuriot, F., Sillon, G., Wenger, A. M., Sureka, D. L., Lester, T., Johnson, D. S., Bowen, J., Calhoun, A. R., Viskochil, D. H., DDD Study, Bejerano, G., Bernstein, J. A., Chitayat, D. Chitayat syndrome: hyperphalangism, characteristic facies, hallux valgus and bronchomalacia results from a recurrent c.266A-G p.(tyr89cys) variant in the ERF gene. J. Med. Genet. 54: 157-165, 2017. [PubMed: 27738187] [Full Text: https://doi.org/10.1136/jmedgenet-2016-104143]
Bose, R., Karthaus, W. R., Armenia, J., Abida, W., Iaquinta, P. J., Zhang, Z., Wongvipat, J., Wasmuth, E. V., Shah, N., Sullivan, P. S., Doran, M. G., Wang, P., Patruno, A., Zhao, Y., International SU2C/PCF Prostate Cancer Dream Team, Zheng, D., Schultz, N., Sawyers, C. L. ERF mutations reveal a balance of ETS factors controlling prostate oncogenesis. Nature 546: 671-675, 2017. [PubMed: 28614298] [Full Text: https://doi.org/10.1038/nature22820]
Chaudhry, A., Sabatini, P., Han, L., Ray, P. N., Forrest, C., Bowdin, S. Heterozygous mutations in ERF cause syndromic craniosynostosis with multiple suture involvement. Am. J. Med. Genet. 167A: 2544-2547, 2015. [PubMed: 26097063] [Full Text: https://doi.org/10.1002/ajmg.a.37218]
Chitayat, D., Haj-Chahine, S., Stalker, H. J., Azouz, E. M., Cote, A., Halal, F. Hyperphalangism, facial anomalies, hallux valgus, and bronchomalacia: a new syndrome? Am. J. Med. Genet. 45: 1-4, 1993. [PubMed: 8418638] [Full Text: https://doi.org/10.1002/ajmg.1320450103]
de Castro, C. M., Rabe, S. M., Langdon, S. D., Fleenor, D. E., Slentz-Kesler, K., Ahmed, M. N., Qumsiyeh, M. B., Kaufman, R. E. Genomic structure and chromosomal localization of the novel ETS factor, PE-2 (ERF). Genomics 42: 227-235, 1997. [PubMed: 9192842] [Full Text: https://doi.org/10.1006/geno.1997.4730]
Janesick, A., Abbey, R., Chung, C., Liu, S., Taketani, M., Blumberg, B. ERF and ETV3L are retinoic acid-inducible repressors required for primary neurogenesis. Development 140: 3095-3106, 2013. [PubMed: 23824578] [Full Text: https://doi.org/10.1242/dev.093716]
Liu, D., Pavlopoulos, E., Modi, W., Moschonas, N., Mavrothalassitis, G. ERF: genomic organization, chromosomal localization and promoter analysis of the human and mouse genes. Oncogene 14: 1445-1451, 1997. [PubMed: 9136988] [Full Text: https://doi.org/10.1038/sj.onc.1200965]
Papadaki, C., Alexiou, M., Cecena, G., Verykokakis, M., Bilitou, A., Cross, J. C., Oshima, R. G., Mavrothalassitis, G. Transcriptional repressor Erf determines extraembryonic ectoderm differentiation. Molec. Cell. Biol. 27: 5201-5213, 2007. [PubMed: 17502352] [Full Text: https://doi.org/10.1128/MCB.02237-06]
Paumard-Hernandez, B., Berges-Soria, J., Barroso, E., Rivera-Pedroza, C. I., Perez-Carrizosa, V., Benito-Sanz, S., Lopez-Messa, E., Santos, F., Garcia-Recuero, I. I., Romance, A., Ballesta-Martinez, M. J., Lopez-Gonzalez, V., Campos-Barros, A., Cruz, J., Guillen-Navarro, E., Sanchez del Pozo, J., Lapunzina, P., Garcia-Minaur, S., Heath, K. E. Expanding the mutation spectrum in 182 Spanish probands with craniosynostosis: identification and characterization of novel TCF12 variants. Europ. J. Hum. Genet. 23: 907-914, 2015. [PubMed: 25271085] [Full Text: https://doi.org/10.1038/ejhg.2014.205]
Sgouras, D. N., Athanasiou, M. A., Beal, G. J., Jr., Fisher, R. J., Blair, D. G., Mavrothalassitis, G. J. ERF: an ETS domain protein with strong transcriptional repressor activity, can suppress ets-associated tumorigenesis and is regulated by phosphorylation during cell cycle and mitogenic stimulation. EMBO J. 14: 4781-4793, 1995. [PubMed: 7588608] [Full Text: https://doi.org/10.1002/j.1460-2075.1995.tb00160.x]
Twigg, S. R. F., Vorgia, E., McGowan, S. J., Peraki, I., Fenwick, A. L., Sharma, V. P., Allegra, M., Zaragkoulias, A., Sadighi Akha, E., Knight, S. J. L., Lord, H., Lester, T., and 15 others. Reduced dosage of ERF causes complex craniosynostosis in humans and mice and links ERF1/2 signaling to regulation of osteogenesis. Nature Genet. 45: 308-313, 2013. [PubMed: 23354439] [Full Text: https://doi.org/10.1038/ng.2539]