PC4 serves as a negative regulator of skin wound healing in mice

Abstract

Background: Human positive cofactor 4 (PC4) was initially characterized as a multifunctional transcriptional cofactor, but its role in skin wound healing is still unclear. The purpose of this study was to explore the role of PC4 in skin wound healing through PC4 knock-in mouse model. Methods: A PC4 knock-in mouse model (PC4+/+) with a dorsal full-thickness wound was used to investigate the biological functions of PC4 in skin wound healing. Quantitative PCR, Western blot analysis and immunohistochemistry were performed to evaluate the expression of PC4; Sirius red staining and immunofluorescence were performed to explore the change of collagen deposition and angiogenesis. Proliferation and apoptosis were detected using Ki67 staining and TUNEL assay. Primary dermal fibroblasts were isolated from mouse skin to perform cell scratch experiments, cck-8 assay and colony formation assay. Results: The PC4+/+ mice were fertile and did not display overt abnormalities but showed an obvious delay in cutaneous healing of dorsal skin. Histological staining showed insufficient re-epithelialization, decreased angiogenesis and collagen deposition, increased apoptosis and decreased cell proliferation in PC4+/+ skin. Our data also showed decreased migration rate and proliferation ability in cultured primary fibroblasts from PC4+/+ mice in vitro. Conclusions: This study suggests that PC4 might serve as a negative regulator of skin wound healing in mice.

Keywords

Wound healing / Skin / PC4 / Proliferation / Migration / Positive cofactor 4

References

[1] Ge H, Roeder RG. Purification, cloning, and characterization of a human coactivator, PC4, that mediates transcriptional activation of class II genes. Cell. 1994; 78: 513-23.

[2] Kretzschmar M, Kaiser K, Lottspeich F, Meisterernst M. A novel mediator of class II gene transcription with homology to viral immediate-early transcriptional regulators. Cell. 1994; 78: 525-34.

[3] Garavís M, Calvo O. Sub1/PC4, a multifaceted factor: from transcription to genome stability. Curr Genet. 2017; 63: 1023-35.

[4] Wang Z, Roeder RG. DNA topoisomerase I and PC4 can interact with human TFIIIC to promote both accurate termination and transcription reinitiation by RNA polymerase III. Mol Cell. 1998; 1: 749-57.

[5] Malik S, Guermah M, Roeder RG. A dynamic model for PC4 coactivator function in RNA polymerase II transcription. Proc Natl Acad Sci USA. 1998; 95: 2192-7.

[6] Calvo O, Manley JL. The transcriptional coactivator PC4/Sub1 has multiple functions in RNA polymerase II transcription. EMBO J. 2005; 24: 1009-20.

[7] Akimoto Y, Yamamoto S, Iida S, Hirose Y, Tanaka A, Hanaoka F, et al. Transcription cofactor PC4 plays essential roles in collaboration with the small subunit of general transcription factor TFIIE. Genes Cells. 2014; 19: 879-90.

[8] Gao J, Zybailov BL, Byrd AK, Griffin WC, Chib S, Mackintosh SG, et al. Yeast transcription co-activator Sub1 and its human homolog PC4 preferentially bind to G-quadruplex DNA. Chem Commun (Camb). 2015; 51: 7242-44.

[9] Huang J, Zhao Y, Liu H, Huang D, Cheng X, Zhao W, et al. Substitution of tryptophan 89 with tyrosine switches the DNA binding mode of PC4. Sci Rep. 2015; 5: 8789.

[10] Mortusewicz O, Evers B, Helleday T. PC4 promotes genome stability and DNA repair through binding of ssDNA at DNA damage sites. Oncogene. 2016; 35: 761-70.

[11] Sancar C, Ha N, Yilmaz R, Tesorero R, Fisher T, Brunner M, et al. Combinatorial control of light induced chromatin remodeling and gene activation in Neurospora. PLoS Genet. 2015; 11: e1005105.

[12] Das C, Hizume K, Batta K, Kumar BR, Gadad SS, Ganguly S, et al. Transcriptional coactivator PC4, a chromatin-associated protein, induces chromatin condensation. Mol Cell Biol. 2006; 26: 8303-15.

[13] Shi C, Mai Y, Zhu Y, Cheng T, Su Y. Spontaneous transformation of a clonal population of dermis-derived multipotent cells in culture. In Vitro Cell Dev Biol Anim. 2007; 43: 290-6.

[14] Peng Y, Yang J, Zhang E, Sun H, Wang Q, Wang T, et al. Human positive coactivator 4 is a potential novel therapeutic target in non-small cell lung cancer. Cancer Gene Ther. 2012; 19: 690-6.

[15] Chen L, Du C, Wang L, Yang C, Zhang JR, Li N, et al. Human positive coactivator 4 (PC4) is involved in the progression and prognosis of astrocytoma. J Neurol Sci. 2014; 346: 293-8.

[16] Luo P, Tan X, Luo S, Wang Z, Long L, Wang Y, et al. An NIR-Fluorophore-based inhibitor of SOD1 induces apoptosis by targeting transcription cofactor PC4. Adv Ther (Weinh). 2019; 2: 1800148.

[17] Luo P, Zhang C, Liao F, Chen L, Liu Z, Long L, et al. Transcriptional positive cofactor 4 promotes breast cancer proliferation and metastasis through c-Myc mediated Warburg effect. Cell Commun Signal. 2019; 17: 36.

[18] Luo P, Jiang Q, Fang Q, Wang Y, Wang Z, Yang J, et al. The human positive cofactor 4 promotes androgen-independent prostate cancer development and progression through HIF-1α/β-catenin pathway. Am J Cancer Res. 2019; 9: 682-98.

[19] Li W, Hou JZ, Niu J, Xi ZQ, Ma C, Sun H, et al. Akt1 inhibition promotes breast cancer metastasis through EGFR-mediated β-catenin nuclear accumulation. Cell Commun Signal. 2018; 16: 82.

[20] Du Z, Luo Q, Yang L, Bing T, Li X, Guo W, et al. Mass spectrometric proteomics reveals that nuclear protein positive cofactor PC4 selectively binds to cross-linked DNA by a trans-platinum anticancer complex. J Am Chem Soc. 2014; 136: 2948-51.

[21] Jo J, Hwang S, Kim HJ, Hong S, Lee JE, Lee SG, et al. An integrated systems biology approach identifies positive cofactor 4 as a factor that increases reprogramming efficiency. Nucleic Acids Res. 2016; 44: 1203-15.

[22] Kurita M, Araoka T, Hishida T, O’Keefe DD, Takahashi Y, Sakamoto A, et al. In vivo reprogramming of wound-resident cells generates skin epithelial tissue. Nature. 2018; 561: 243-7.

[23] Eming SA, Martin P, Tomic-Canic M. Wound repair and regeneration: mechanisms, signaling, and translation. Sci Transl Med. 2014; 6: 265sr6.

[24] Dekoninck S, Blanpain C. Stem cell dynamics, migration and plasticity during wound healing. Nat Cell Biol. 2019; 21: 18-24.

[25] Miura T, Kawakami K, Kanno E, Tanno H, Tada H, Sato N, et al. Dectin-2-mediated Signaling leads to delayed skin wound healing through enhanced Neutrophilic inflammatory response and neutrophil extracellular trap formation. J Invest Dermatol. 2019; 139: 702-11.

[26] Liang G, Fu W, Wang K. Analysis of t-test misuses and SPSS operations in medical research papers. Burns Trauma. 2019; 7: 31.

[27] Ma T, Fu B, Yang X, Xiao Y, Pan M. Adipose mesenchymal stem cell-derived exosomes promote cell proliferation, migration, and inhibit cell apoptosis via Wnt/β-catenin signaling in cutaneous wound healing. J Cell Biochem. 2019; 120: 10847-54.

[28] Justet C, Hernández JA, Torriglia A, Chifflet S. Fast calcium wave inhibits excessive apoptosis during epithelial wound healing. Cell Tissue Res. 2016; 365: 343-56.

[29] Duscher D, Maan ZN, Whittam AJ, Sorkin M, Hu MS, Walmsley GG, et al. Fibroblast-specific deletion of hypoxia inducible Factor-1 critically impairs murine cutaneous neovascularization and wound healing. Plast Reconstr Surg. 2015; 136: 1004-13.

[30] Icli B, Nabzdyk CS, Lujan-Hernandez J, Cahill M, Auster ME, Wara AK, et al. Regulation of impaired angiogenesis in diabetic dermal wound healing by microRNA-26a. J Mol Cell Cardiol. 2016; 91: 151-9.

[31] Rodrigues M, Kosaric N, Bonham CA, Gurtner GC. Wound healing: a cellular perspective. Physiol Rev. 2019; 99: 665-706.

[32] Iacopetti P, Barsacchi G, Tirone F, Cremisi F. Expression of the PC4 gene in the developing rat nervous system. Brain Res. 1996; 707: 293-7.

[33] Pröls F, Loser B, Marx M. Differential expression of osteopontin, PC4, and CEC5, a novel mRNA species, during in vitro angiogenesis. Exp Cell Res. 1998; 239: 1-10.

[34] Malik S, Guermah M, Roeder RG. A dynamic model for PC4 coactivator function in RNA polymerase II transcription. Proc Natl Acad Sci USA. 1998; 95: 2192-7.

[35] Tavenet A, Suleau A, Dubreuil G, Ferrari R, Ducrot C, Michaut M, et al. Genome-wide location analysis reveals a role for Sub1 in RNA polymerase III transcription. Proc Natl Acad Sci USA. 2009; 106: 14265-70.

[36] Das C, Gadad SS, Kundu TK. Human positive coactivator 4 controls heterochromatinization and silencing of neural gene expression by interacting with REST/NRSF and CoREST. J Mol Biol. 2010; 397: 1-12.