A Determination of Valosin-Containing Protein (p97/VCP) and Small VCP-Interacting Protein (SVIP) Expressions in Rat Kidney and Vero Cells
DOI:
https://doi.org/10.58600/eurjther3030Keywords:
endoplasmic reticulum-associated protein degradation (ERAD), kidney, p97/valosin containing protein (VCP), rat, small VCP-interacting protein (SVIP), vero cellsAbstract
Objective: The 97-kDa valosin-containing protein (p97/VCP), which is involved in the ubiquitin-proteasome-mediated degradation pathway, regulates a diversity of cellular activities. p97/VCP, which also physically interacts with the 26 proteasome and uses multiubiquitin chains to immediately attach the ubiquitinated substrates. Small p97/VCP-interacting protein (SVIP), was first detected to be one of numerous cofactors controlling the VCP. In rat kidney tissues and Vero cells, SVIP and p97/VCP proteins haven’t been investigated. The aim of present study was to examine the expression of p97/VCP and SVIP in kidney tissues and Vero cells. Also, this study was purposed to conduce to further research on the use of two proteins as kidney cell biomarkers in cases of renal pathologies.
Methods: Hematoxylin-eosin (HE) staining was used to examine the histomorphology of rat kidney tissues. The cellular distribution and expression of SVIP and p97/VCP in rat renal tissue were investigated using the immunohistochemistry method. Also, Vero kidney cell line was used to determine cellular localization and expression of ERAD proteins by immunofluorescence and Western blotting.
Results: Immunohistochemistry analysis revealed that SVIP and p97/VCP were primarily located in the nucleus and cytoplasm of the proximal and distal tubule cells in the rat kidney. Furthermore, the immunoexpression of these proteins was discovered in the epithelial cells of the Henle loop and collecting tubules in the medulla. In the nucleus and cytoplasm of Vero kidney cells in vitro, p97/VCP immunoexpression was detected. SVIP was determined in the cytoplasm of the cells. Also, Vero cells showed intense p97/VCP expression compared to SVIP expression by Western blotting.
Conclusions: In conclusion, the current study showed that p97/VCP and SVIP had cellular localization and protein expression in the rat kidney tissue and kidney cell line (Vero). We might advise that additional research is required to ascertain the role of SVIP and p97/VCP in renal tissue.
References
[1] Xia D, Tang WK, Ye Y (2016) Structure and function of the AAA+ ATPase p97/Cdc48p. Gene. 583(1):64–77. https://doi.org/10.1016/j.gene.2016.02.042
[2] Buchberger A, Schindelin H, Hanzelmann P (2015) Control of p97 function by cofactor binding. FEBS Lett. 589:2578–2589. https://doi.org/10.1016/j.febslet.2015.08.028
[3] Nandi D, Tahiliani P, Kumar A, Chandu D (2006) The ubiquitin-proteasome system. J Biosci. 31:137–155. https://doi.org/10.1007/BF02705243
[4] Kleiger G, Mayor T (2014) Perilous journey: A tour of the ubiquitin-proteasome system. Trends Cell Biol. 24:352–359. https://doi.org/10.1016/j.tcb.2013.12.003
[5] Kwon YT, Ciechanover A (2017) The ubiquitin code in the ubiquitin-proteasome system and autophagy. Trends Biochem Sci. 42:873–886. https://doi.org/10.1016/j.tibs.2017.09.002
[6] Glickman MH, Ciechanover A (2002) The ubiquitin-proteasome proteolytic pathway: Destruction for the sake of construction. Physiol Rev. 82:373–428. https://doi.org/10.1152/physrev.00027.2001
[7] Meyer H, Bug M, Bremer S (2012) Emerging functions of the VCP/p97 AAA-ATPase in the ubiquitin system. Nat Cell Biol. 14:117–123. https://doi.org/10.1038/ncb2407
[8] Van den Boom J, Meyer H (2018) VCP/p97-mediated unfolding as a principle in protein homeostasis and signaling. Mol Cell. 69(2):182–194. https://doi.org/10.1016/j.molcel.2017.10.028
[9] Costantini S, Capone F, Polo A, Bagnara P, Budillon A (2021) Valosin-containing protein (VCP)/p97: A prognostic biomarker and therapeutic target in cancer. Int J Mol Sci. 22:10177. https://doi.org/10.3390/ijms221810177
[10] Yamanaka K, Sasagawa Y, Ogura T (2012) Recent advances in p97/VCP/Cdc48 cellular functions. Biochim Biophys Acta. 1823:130–137. https://doi.org/10.1016/j.bbamcr.2011.07.001
[11] Ye Y, Tang WK, Zhang T, Xia D (2017) A mighty “protein extractor” of the cell: Structure and function of the p97/CDC48 ATPase. Front Mol Biosci. 4:39. https://doi.org/10.3389/fmolb.2017.00039
[12] Tsai YC, Weissman AM (2010) The unfolded protein response, degradation from endoplasmic reticulum and cancer. Genes Cancer. 7:764–778. https://doi.org/10.1177/1947601910383011.
[13] Segura-Cabrera A, Tripathi R, Zhang X, Gui L, Chou TF, Komurov K (2017) A structure- and chemical genomics-based approach for repositioning of drugs against VCP/p97 ATPase. Sci Rep. 7:44912. https://doi.org/10.1038/srep44912
[14] Bento AC, Bippes CC, Kohler C, Hemion C, Frank S, Neutzner A (2018) UBXD1 is a mitochondrial recruitment factor for p97/VCP and promotes mitophagy. Sci Rep. 8:12415. https://doi.org/10.1038/s41598-018-30963-z
[15] Mengus C, Neutzner M, Bento A, Bippes CC, Kohler C, Decembrini S, Häusel J, Hemion C, Sironi L, Frank S, Scholl HPN, Neutzner A (2022) VCP/p97 cofactor UBXN1/SAKS1 regulates mitophagy by modulating MFN2 removal from mitochondria. Autophagy. 18:171–190. https://doi.org/10.1080/15548627.2021.1922982
[16] Ballar P, Fang S (2008) Regulation of ER-associated degradation via p97/VCP-interacting motif. Biochem Soc Trans. 36:818–822. https://doi.org/10.1042/BST0360818
[17] Ballar P, Zhong Y, Nagahama M, Tagaya M, Shen Y, Fang S (2007) Identification of SVIP as an endogenous inhibitor of ER-associated degradation. J. Biol. Chem. 282,33908–33914. https://doi.org/10.1074/jbc.M704446200
[18] Zou HM, Yu J, Ruan YY, Xie Y, An XM, Chen PL, Luo YQ, Shi MJ, Liu M, Xu LF, Liu J, Guo B, Zhang F (2024) HNF-1β alleviates podocyte injury in lupus nephritis by maintaining endoplasmic reticulum homeostasis. Lupus science & medicine. 11(2), e001349. https://doi.org/10.1136/lupus-2024-001349
[19] Huang Z, Hong Q, Xue P, Paul G, Feng Z, Wang L, Mei Y, Wu L, Chen X, Wu D (2012) A proteome-wide screen identifies valosin-containing protein as an essential regulator of podocyte endoplasmic reticulum stress. Chin. Sci. Bull. 57,2493–2505 https://doi.org/10.1007/s11434-012-5250-8.
[20] Akcan G, Alimogullari E, Abu-Issa R, Cayli S (2020) Analysis of the developmental expression of small VCP-interacting protein and its interaction with steroidogenic acute regulatory protein in Leydig cells. Reprod Biol. 20:88–96. https://doi.org/10.1016/j.repbio.2020.01.006
[21] Wang Y, Ballar P, Zhong Y, Zhang X, Liu C, Zhang YJ, Monteiro MJ, Li J, Fang S (2011) SVIP induces localization of p97/VCP to the plasma and lysosomal membranes and regulates autophagy. PLoS One. 6:e24478. https://doi.org/10.1371/journal.pone.0024478
[22] Jia D, Wang YY, Wang P, Huang Y, Liang DY, Wang D, Cheng C, Zhang C, Guo L, Liang P, Wang Y, Jia Y, Li C (2019) SVIP alleviates CCl4-induced liver fibrosis via activating autophagy and protecting hepatocytes. Cell Death Dis. 10:71. https://doi.org/10.1038/s41419-019-1311-0
[23] Fujimoto D, Kuwabara T, Hata Y, Umemoto S, Kanki T, Nishiguchi Y, Mizumoto T, Hayata M, Kakizoe Y, Izumi Y, Takahashi S, Mukoyama M (2020) Suppressed ER-associated degradation by intraglomerular cross talk between mesangial cells and podocytes causes podocyte injury in diabetic kidney disease. FASEB J. 34:15577–15590. https://doi.org/10.1096/fj.202000078RR
[24] Yoshida S, Wei X, Zhang G, O'Connor CL, Torres M, Zhou Z, Lin L, Menon R, Xu X, Zheng W, Xiong Y, Otto E, Tang CA, Hua R, Verma R, Mori H, Zhang Y, Hu CA, Liu M, Garg P, Hodgin JB, Sun S, Bitzer M, Qi L (2021) Endoplasmic reticulum-associated degradation is required for nephrin maturation and kidney glomerular filtration function. J Clin Invest. 131:e143988. https://doi.org/10.1172/JCI143988
[25] Ren G, Tardi NJ, Matsuda F, Koh KH, Ruiz P, Wei C, Altintas MM, Ploegh H, Reiser J (2018) Podocytes exhibit a specialized protein quality control employing derlin-2 in kidney disease. Am J Physiol Renal Physiol. 314:F471–F482. https://doi.org/10.1152/ajprenal.00691.2016
[26] Nezvitsky L, Tremblay ML, Takano T, Papillon J, Cybulsky AV (2014) Complement-mediated glomerular injury is reduced by inhibition of protein-tyrosine phosphatase 1B. Am J Physiol Renal Physiol. 307:F634–647. https://doi.org/10.1152/ajprenal.00191.2014
[27] Cayli S, Erdemir F, Ocaklı S, Ungor B, Kesici H, Yener T, Aslan H (2012) Interaction between Smad1 and p97/VCP in rat testis and epididymis during postnatal development. Reprod Sci. 19:190–201. https://doi.org/10.1177/1933719111417886
[28] Moir D, Stewart SE, Osmond BC, Botstein D (1982) Cold-sensitive cell-division-cycle mutants of yeast: Isolation, properties, and pseudoreversion studies. Genetics. 100:547–563. https://doi.org/10.1093/genetics/100.4.547
[29] Dai RM, Li CC (2001) Valosin-containing protein is a multi-ubiquitin chain-targeting factor required in ubiquitin-proteasome degradation. Nat Cell Biol. 3:740–744. https://doi.org/10.1038/35087056
[30] Christianson JC, Ye Y (2014) Cleaning up in the endoplasmic reticulum: Ubiquitin in charge. Nat Struct Mol Biol. 21:325–335. https://doi.org/10.1038/nsmb.2793
[31] Ruggiano A, Foresti O, Carvalho P (2014) Quality control: ER-associated degradation: Protein quality control and beyond. J Cell Biol. 204:869–879. https://doi.org/10.1083/jcb.201312042
[32] Smith MH, Ploegh HL, Weissman JS (2011) Road to ruin: Targeting proteins for degradation in the endoplasmic reticulum. Science. 334:1086–1090. https://doi.org/10.1126/science.1209235
[33] Carvalho P, Stanley AM, Rapoport TA (2010) Retrotranslocation of a misfolded luminal ER protein by the ubiquitin-ligase Hrd1p. Cell. 143:579–591. https://doi.org/10.1016/j.cell.2010.10.028
[34] Beskow A, Grimberg KB, Bott LC, Salomons FA, Dantuma NP, Young P (2009) A conserved unfoldase activity for the p97 AAA-ATPase in proteasomal degradation. J Mol Biol. 394:732–746. https://doi.org/10.1016/j.jmb.2009.09.050
[35] Gallagher PS, Clowes Candadai SV, Gardner RG (2014) The requirement for Cdc48/p97 in nuclear protein quality control degradation depends on the substrate and correlates with substrate insolubility. J Cell Sci. 127:1980–1991. https://doi.org/10.1242/jcs.141838
[36] Qu J, Zou T, Lin Z (2021) The roles of the ubiquitin–proteasome system in the endoplasmic reticulum stress pathway. Int J Mol Sci. 22(4):1526. https://doi.org/10.3390/ijms22041526
[37] Vaz B, Halder S, Ramadan K (2013) Role of p97/VCP (Cdc48) in genome stability. Front Genet. 4:60. https://doi.org/10.3389/fgene.2013.00060
[38] Meyer HH (2005) Golgi reassembly after mitosis: The AAA family meets the ubiquitin family. Biochim Biophys Acta. 1744:108–119. https://doi.org/10.1016/j.bbamcr.2005.03.011
[39] Olzmann JA, Richter CM, Kopito RR (2013) Spatial regulation of UBXD8 and p97/VCP controls ATGL-mediated lipid droplet turnover. Proc Natl Acad Sci USA. 110:1345–1350. https://doi.org/10.1073/pnas.1213738110
[40] Meyer H, Weihl CC (2014) The VCP/p97 system at a glance: Connecting cellular function to disease pathogenesis. J Cell Sci. 127:3877–3883. https://doi.org/10.1242/jcs.093831
[41] Ortak H, Cayli S, Tas U, Ocakli S, Söğüt E, Demir HD (2012) Expression of p97/VCP and ubiquitin during postnatal development of the degenerating rat retina. J Mol Histol. 43:17–25. https://doi.org/10.1007/s10735-011-9374-y
[42] Wertz K, Herrmann BG (2000) Large-scale screen for genes involved in gonad development. Mech Dev. 98:51–70. https://doi.org/10.1016/S0925-4773(00)00452-4
[43] Nakatsuji T (2013) Clinical features of renal cell carcinoma (RCC) found in 110 nephrectomized Japanese, of which 24 (22%) RCC cases showed 31 double or triple cancers. Comp Clin Pathol. 22, 367–378. https://doi.org/10.1007/s00580-012-1418-5
[44] Nagahama M, Suzuki M, Hamada Y, Hatsuzawa K, Tani K, Yamamoto A, Tagaya M (2003) SVIP is a novel VCP/p97-interacting protein whose expression causes cell vacuolation. Mol Biol Cell. 14:262–273. https://doi.org/10.1091/mbc.02-07-0115
[45] Romanuik TL, Wang G, Holt RA, Jones SJM, Marra MA, Sadar MD (2009) Identification of novel androgen-responsive genes by sequencing of longSAGE libraries. BMC Genomics. 10:476. https://doi.org/10.1186/1471-2164-10-476
[46] Erzurumlu Y, Ballar P (2016) Androgen mediated regulation of endoplasmic reticulum-associated degradation and its effects on prostate cancer. Sci Rep. 7:40719. https://doi.org/10.1038/srep40719
[47] Johnson AE, Orr BO, Fetter RD, Moughamian AJ, Primeaux LA, Geier EG, Yokoyama JS, Miller BL, Davis GW (2021) SVIP is a molecular determinant of lysosomal dynamic stability, neurodegeneration, and lifespan. Nat Commun. 1:513. https://doi.org/10.1038/s41467-020-20796-8
[48] Ilhan R, Üner G, Yilmaz S, Atalay Sahar E, Cayli S, Erzurumlu Y, Gozen O, Kirmizibayrak PB (2022) Novel regulation mechanism of adrenal cortisol and DHEA biosynthesis via the endogenous ERAD inhibitor small VCP-interacting protein. Sci Rep. 12:869. https://doi.org/10.1038/s41598-022-04821-y
[49] Cayli S, Alimogullari E, Piskin I, Bilginoglu A, Nakkas H (2021) Effect of pioglitazone on the expression of ubiquitin proteasome system and autophagic proteins in rat pancreas with metabolic syndrome. J Mol Histol. 52(5):929-942. https://doi.org/10.1007/s10735-021-10013-1
[50] Woźniak MJ, Bola B, Brownhill K, Yang YC, Levakova V, Allan VJ (2009) Role of kinesin-1 and cytoplasmic dynein in endoplasmic reticulum movement in VERO cells. J Cell Sci. 122(Pt 12):1979–1989. https://doi.org/10.1242/jcs.041962
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