A Method for In Situ Reverse Genetic Analysis of Proteins Involved mtDNA Replication.
GeneSwap approach
TFAM
TFAM chimeras
TFAM knockout
TFAM-mtDNA evolutionary co-adaptation
mtDNA instability
mtDNA metabolism
mtDNA replication
mtDNA transcription
Journal
Cells
ISSN: 2073-4409
Titre abrégé: Cells
Pays: Switzerland
ID NLM: 101600052
Informations de publication
Date de publication:
11 07 2022
11 07 2022
Historique:
received:
09
06
2022
revised:
29
06
2022
accepted:
10
07
2022
entrez:
27
7
2022
pubmed:
28
7
2022
medline:
29
7
2022
Statut:
epublish
Résumé
The unavailability of tractable reverse genetic analysis approaches represents an obstacle to a better understanding of mitochondrial DNA replication. Here, we used CRISPR-Cas9 mediated gene editing to establish the conditional viability of knockouts in the key proteins involved in mtDNA replication. This observation prompted us to develop a set of tools for reverse genetic analysis in situ, which we called the GeneSwap approach. The technique was validated by identifying 730 amino acid (aa) substitutions in the mature human TFAM that are conditionally permissive for mtDNA replication. We established that HMG domains of TFAM are functionally independent, which opens opportunities for engineering chimeric TFAMs with customized properties for studies on mtDNA replication, mitochondrial transcription, and respiratory chain function. Finally, we present evidence that the HMG2 domain plays the leading role in TFAM species-specificity, thus indicating a potential pathway for TFAM-mtDNA evolutionary co-adaptations.
Identifiants
pubmed: 35883613
pii: cells11142168
doi: 10.3390/cells11142168
pmc: PMC9316749
pii:
doi:
Substances chimiques
DNA, Mitochondrial
0
DNA-Binding Proteins
0
Mitochondrial Proteins
0
Transcription Factors
0
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, U.S. Gov't, Non-P.H.S.
Langues
eng
Sous-ensembles de citation
IM
Subventions
Organisme : NIH HHS
ID : S10 OD025089
Pays : United States
Organisme : NIH HHS
ID : R01 OD010944
Pays : United States
Organisme : NHLBI NIH HHS
ID : P01 HL066299
Pays : United States
Références
Mol Cell Biol. 1988 Aug;8(8):3496-509
pubmed: 3211148
Cell. 1983 Aug;34(1):151-9
pubmed: 6883508
J Biol Chem. 2003 Aug 15;278(33):31149-58
pubmed: 12759347
PLoS One. 2018 Mar 28;13(3):e0194887
pubmed: 29590189
Exp Mol Med. 2015 Mar 13;47:e150
pubmed: 25766619
Mol Genet Metab. 2016 Sep;119(1-2):91-9
pubmed: 27448789
Proc Natl Acad Sci U S A. 2012 Oct 9;109(41):16510-5
pubmed: 23012404
Proc Natl Acad Sci U S A. 2012 Apr 24;109(17):6513-8
pubmed: 22493245
Life (Basel). 2021 Dec 24;12(1):
pubmed: 35054416
PLoS One. 2020 Jul 10;15(7):e0235856
pubmed: 32649732
Cell Metab. 2015 Sep 1;22(3):485-98
pubmed: 26299452
Hum Mol Genet. 2013 Mar 1;22(5):1017-25
pubmed: 23197651
Mitochondrion. 2020 Jul;53:133-139
pubmed: 32470614
Transcription. 2011 Mar;2(2):55-59
pubmed: 21468229
Biochem Biophys Res Commun. 2014 Jul 18;450(1):166-71
pubmed: 24875355
Mol Biol Cell. 2001 Apr;12(4):821-30
pubmed: 11294889
Hum Mol Genet. 2005 Jul 1;14(13):1775-83
pubmed: 15888483
Proc Natl Acad Sci U S A. 2004 Mar 2;101(9):3136-41
pubmed: 14978272
Proc Natl Acad Sci U S A. 2010 Jul 6;107(27):12133-8
pubmed: 20562347
Cell. 1987 Jul 17;50(2):247-58
pubmed: 3594571
Mol Cell Biol. 2004 Nov;24(22):9823-34
pubmed: 15509786
Proc Natl Acad Sci U S A. 1997 Aug 19;94(17):9131-5
pubmed: 9256447
Science. 2015 Nov 27;350(6264):1092-6
pubmed: 26472760
Science. 2015 Nov 27;350(6264):1096-101
pubmed: 26472758
Proc Natl Acad Sci U S A. 2012 Apr 24;109(17):6508-12
pubmed: 22454497
Annu Rev Genet. 2005;39:359-407
pubmed: 16285865
Gene. 1989 Apr 15;77(1):51-9
pubmed: 2744487
Nucleic Acids Res. 2010 Oct;38(19):6577-88
pubmed: 20566479
Cell Metab. 2013 Apr 2;17(4):618-26
pubmed: 23562081
J Biol Chem. 2013 Sep 13;288(37):26594-605
pubmed: 23884459
Cell. 2005 Dec 29;123(7):1227-40
pubmed: 16377564
DNA Repair (Amst). 2015 Dec;36:122-136
pubmed: 26411874
FEBS J. 2007 Dec;274(24):6488-99
pubmed: 18028422
Front Biosci (Landmark Ed). 2017 Jan 1;22(5):835-853
pubmed: 27814650
Biochim Biophys Acta Mol Cell Res. 2022 Jan;1869(1):119167
pubmed: 34744028
Cardiovasc Res. 2011 Apr 1;90(1):57-67
pubmed: 21113058
Hum Genet. 2021 Dec;140(12):1733-1751
pubmed: 34647195
PLoS One. 2019 Mar 7;14(3):e0213376
pubmed: 30845180
Genome Med. 2020 Sep 28;12(1):84
pubmed: 32988399
J Genet Genomics. 2009 Mar;36(3):125-31
pubmed: 19302968
Biochim Biophys Acta. 2009 Dec;1792(12):1103-8
pubmed: 19596444
Annu Rev Biochem. 2016 Jun 2;85:133-60
pubmed: 27023847
Nat Genet. 1998 Mar;18(3):231-6
pubmed: 9500544
Sci Adv. 2016 Aug 05;2(8):e1600963
pubmed: 27532055
Front Biosci (Landmark Ed). 2014 Jan 01;19(5):777-82
pubmed: 24389221
Nat Biotechnol. 2016 Jun;34(6):634-6
pubmed: 27159373
Curr Protoc Cell Biol. 2018 Mar;78(1):20.11.1-20.11.14
pubmed: 30040188
Science. 2013 Feb 15;339(6121):819-23
pubmed: 23287718
PLoS One. 2015 Apr 24;10(4):e0124633
pubmed: 25909470
Curr Pharm Des. 2015;21(9):1158-63
pubmed: 25312735
Stem Cells. 2011 Sep;29(9):1459-68
pubmed: 21780252
Science. 2020 Jun 19;368(6497):1371-1376
pubmed: 32439659
Mitochondrion. 2021 Nov;61:102-113
pubmed: 34606994
Nat Metab. 2019 Dec;1(12):1209-1218
pubmed: 32395698
Nat Rev Cancer. 2012 Oct;12(10):685-98
pubmed: 23001348
Nat Methods. 2012 Oct;9(10):973-5
pubmed: 22941364
Proc Natl Acad Sci U S A. 1982 Dec;79(23):7195-9
pubmed: 6185947