Forward genetic screen in human podocytes identifies diphthamide biosynthesis genes as regulators of adhesion.
Animals
Cell Adhesion
/ genetics
Cell Line
Drosophila
Fibronectins
/ metabolism
HSP40 Heat-Shock Proteins
/ genetics
High-Throughput Screening Assays
Histidine
/ analogs & derivatives
Humans
Intracellular Signaling Peptides and Proteins
/ genetics
Kidney
/ metabolism
Minor Histocompatibility Antigens
/ genetics
Podocytes
/ metabolism
Proteins
/ genetics
RNA, Small Interfering
Tumor Suppressor Proteins
/ genetics
Vascular Endothelial Growth Factor Receptor-1
/ genetics
adhesion
diphthamide
genetic screen
glomerulus
podocyte
Journal
American journal of physiology. Renal physiology
ISSN: 1522-1466
Titre abrégé: Am J Physiol Renal Physiol
Pays: United States
ID NLM: 100901990
Informations de publication
Date de publication:
01 12 2019
01 12 2019
Historique:
pubmed:
1
10
2019
medline:
14
4
2020
entrez:
1
10
2019
Statut:
ppublish
Résumé
Podocyte function is tightly linked to the complex organization of its cytoskeleton and adhesion to the underlying glomerular basement membrane. Adhesion of cultured podocytes to a variety of substrates is reported to correlate with podocyte health. To identify novel genes that are important for podocyte function, we designed an in vitro genetic screen based on podocyte adhesion to plates coated with either fibronectin or soluble Fms-like tyrosine kinase-1 (sFLT1)/Fc. A genome-scale pooled RNA interference screen on immortalized human podocytes identified 77 genes that increased adhesion to fibronectin, 101 genes that increased adhesion to sFLT1/Fc, and 44 genes that increased adhesion to both substrates when knocked down. Multiple shRNAs against diphthamide biosynthesis protein 1-4 (DPH1-DPH4) were top hits for increased adhesion. Immortalized human podocyte cells stably expressing these hairpins displayed increased adhesion to both substrates. We then used CRISPR-Cas9 to generate podocyte knockout cells for
Identifiants
pubmed: 31566424
doi: 10.1152/ajprenal.00195.2019
pmc: PMC6962514
doi:
Substances chimiques
DNAJC24 protein, human
0
DPH1 protein, human
0
DPH2 protein, human
0
DPH3 protein, human
0
Fibronectins
0
HSP40 Heat-Shock Proteins
0
Intracellular Signaling Peptides and Proteins
0
Minor Histocompatibility Antigens
0
Proteins
0
RNA, Small Interfering
0
Tumor Suppressor Proteins
0
Histidine
4QD397987E
diphthamide
75645-22-6
FLT1 protein, human
EC 2.7.10.1
Vascular Endothelial Growth Factor Receptor-1
EC 2.7.10.1
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
F1593-F1604Subventions
Organisme : Medical Research Council
ID : G0800200
Pays : United Kingdom
Organisme : Medical Research Council
ID : MR/P024297/1
Pays : United Kingdom
Organisme : Medical Research Council
ID : MR/L002418/1
Pays : United Kingdom
Organisme : NEI NIH HHS
ID : R01 EY025799
Pays : United States
Organisme : CIHR
ID : M0P77756
Pays : Canada
Organisme : Medical Research Council
ID : G0800571
Pays : United Kingdom
Organisme : NHLBI NIH HHS
ID : R01 HL124120
Pays : United States
Organisme : NIDDK NIH HHS
ID : R01 DK098410
Pays : United States
Organisme : CIHR
ID : M0P62931
Pays : Canada
Organisme : Medical Research Council
ID : MR/R013942/1
Pays : United Kingdom
Organisme : Medical Research Council
ID : MR/R003017/1
Pays : United Kingdom
Références
Dev Biol. 2008 Apr 15;316(2):288-301
pubmed: 18328474
Crit Rev Biochem Mol Biol. 2013 Nov-Dec;48(6):515-21
pubmed: 23971743
J Am Soc Nephrol. 2017 Nov;28(11):3166-3174
pubmed: 28864466
Proc Natl Acad Sci U S A. 2012 Aug 21;109(34):13817-22
pubmed: 22869748
J Nutr Biochem. 2015 Feb;26(2):120-9
pubmed: 25465156
J Am Soc Nephrol. 2010 Apr;21(4):579-86
pubmed: 20150535
Am J Physiol Renal Physiol. 2008 Nov;295(5):F1535-44
pubmed: 18784258
Nat Rev Genet. 2006 May;7(5):373-84
pubmed: 16607398
J Am Soc Nephrol. 2008 Apr;19(4):677-84
pubmed: 18235087
Kidney Int. 2014 Jan;85(1):124-33
pubmed: 24048372
Acta Biochim Biophys Sin (Shanghai). 2012 Feb;44(2):103-12
pubmed: 22271906
J Clin Invest. 2014 Mar;124(3):1098-113
pubmed: 24531545
J Clin Invest. 2013 Dec;123(12):5179-89
pubmed: 24270420
J Am Soc Nephrol. 2006 May;17(5):1334-44
pubmed: 16611717
Nucleic Acids Res. 2017 Jan 4;45(D1):D663-D671
pubmed: 27799470
PLoS One. 2012;7(11):e49988
pubmed: 23185508
BMC Genomics. 2011 May 06;12:213
pubmed: 21548937
J Clin Invest. 2011 Jun;121(6):2181-96
pubmed: 21606597
Exp Cell Res. 2004 Mar 10;294(1):68-76
pubmed: 14980502
BMC Nephrol. 2008 Jul 22;9:7
pubmed: 18647412
Nat Cell Biol. 2015 Dec;17(12):1577-1587
pubmed: 26479319
J Am Soc Nephrol. 2005 Jul;16(7):1966-76
pubmed: 15872073
J Am Soc Nephrol. 2007 Nov;18(11):3004-16
pubmed: 17942953
Front Endocrinol (Lausanne). 2014 Oct 14;5:160
pubmed: 25352829
BMC Bioinformatics. 2011 Aug 31;12:357
pubmed: 21880147
Kidney Int. 2011 Nov;80(10):1045-54
pubmed: 21814175
Nat Rev Nephrol. 2013 Apr;9(4):200-10
pubmed: 23338211
J Am Soc Nephrol. 1999 Aug;10(8):1633-9
pubmed: 10446930
N Engl J Med. 2012 Apr 19;366(16):1508-14
pubmed: 22512483
Genes Dev. 2004 Feb 1;18(3):320-32
pubmed: 14744934
Redox Biol. 2019 Jan;20:146-156
pubmed: 30312900
Hum Mol Genet. 2017 Feb 15;26(4):768-780
pubmed: 28164240
J Am Soc Nephrol. 2013 Feb;24(2):191-7
pubmed: 23291470
FEBS Lett. 2002 Dec 4;532(1-2):211-5
pubmed: 12459492
J Am Soc Nephrol. 2006 Dec;17(12):3438-46
pubmed: 17082236
Nat Genet. 1995 Aug;10(4):400-6
pubmed: 7670489
Cell. 2012 Oct 12;151(2):384-99
pubmed: 23063127
Genes Chromosomes Cancer. 2008 Apr;47(4):315-25
pubmed: 18181179
J Am Soc Nephrol. 2013 Feb;24(2):161-3
pubmed: 23334393
Hum Mol Genet. 2004 Nov 1;13(21):2625-32
pubmed: 15367484
Nat Rev Mol Cell Biol. 2014 Apr;15(4):273-88
pubmed: 24651544
Genome Biol. 2006;7(10):R100
pubmed: 17076895
Nat Med. 2011 Jul 31;17(8):952-60
pubmed: 21804539