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Physical and functional interactions of a monothiol glutaredoxin and an iron sulfur cluster carrier protein with the sulfur-donating radical S-adenosyl-L-methionine enzyme MiaB.
J Biol Chem 2013; 288(20):14200-11JB

Abstract

The biosynthesis of iron sulfur (FeS) clusters, their trafficking from initial assembly on scaffold proteins via carrier proteins to final incorporation into FeS apoproteins, is a highly coordinated process enabled by multiprotein systems encoded in iscRSUAhscBAfdx and sufABCDSE operons in Escherichia coli. Although these systems are believed to encode all factors required for initial cluster assembly and transfer to FeS carrier proteins, accessory factors such as monothiol glutaredoxin, GrxD, and the FeS carrier protein NfuA are located outside of these defined systems. These factors have been suggested to function both as shuttle proteins acting to transfer clusters between scaffold and carrier proteins and in the final stages of FeS protein assembly by transferring clusters to client FeS apoproteins. Here we implicate both of these factors in client protein interactions. We demonstrate specific interactions between GrxD, NfuA, and the methylthiolase MiaB, a radical S-adenosyl-L-methionine-dependent enzyme involved in the maturation of a subset of tRNAs. We show that GrxD and NfuA physically interact with MiaB with affinities compatible with an in vivo function. We furthermore demonstrate that NfuA is able to transfer its cluster in vitro to MiaB, whereas GrxD is unable to do so. The relevance of these interactions was demonstrated by linking the activity of MiaB with GrxD and NfuA in vivo. We observe a severe defect in in vivo MiaB activity in cells lacking both GrxD and NfuA, suggesting that these proteins could play complementary roles in maturation and repair of MiaB.

Authors+Show Affiliations

Life Sciences Division, Lawrence Berkeley National Laboratory, Berkeley, California 94720, USA.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

Language

eng

PubMed ID

23543739

Citation

Boutigny, Sylvain, et al. "Physical and Functional Interactions of a Monothiol Glutaredoxin and an Iron Sulfur Cluster Carrier Protein With the Sulfur-donating Radical S-adenosyl-L-methionine Enzyme MiaB." The Journal of Biological Chemistry, vol. 288, no. 20, 2013, pp. 14200-11.
Boutigny S, Saini A, Baidoo EE, et al. Physical and functional interactions of a monothiol glutaredoxin and an iron sulfur cluster carrier protein with the sulfur-donating radical S-adenosyl-L-methionine enzyme MiaB. J Biol Chem. 2013;288(20):14200-11.
Boutigny, S., Saini, A., Baidoo, E. E., Yeung, N., Keasling, J. D., & Butland, G. (2013). Physical and functional interactions of a monothiol glutaredoxin and an iron sulfur cluster carrier protein with the sulfur-donating radical S-adenosyl-L-methionine enzyme MiaB. The Journal of Biological Chemistry, 288(20), pp. 14200-11. doi:10.1074/jbc.M113.460360.
Boutigny S, et al. Physical and Functional Interactions of a Monothiol Glutaredoxin and an Iron Sulfur Cluster Carrier Protein With the Sulfur-donating Radical S-adenosyl-L-methionine Enzyme MiaB. J Biol Chem. 2013 May 17;288(20):14200-11. PubMed PMID: 23543739.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Physical and functional interactions of a monothiol glutaredoxin and an iron sulfur cluster carrier protein with the sulfur-donating radical S-adenosyl-L-methionine enzyme MiaB. AU - Boutigny,Sylvain, AU - Saini,Avneesh, AU - Baidoo,Edward E K, AU - Yeung,Natasha, AU - Keasling,Jay D, AU - Butland,Gareth, Y1 - 2013/03/29/ PY - 2013/4/2/entrez PY - 2013/4/2/pubmed PY - 2013/8/7/medline KW - Carrier Protein KW - Escherichia coli KW - Gene Knockout KW - Iron Sulfur Cluster Biosynthesis KW - Iron-Sulfur Protein KW - Monothiol Glutaredoxin KW - Protein-Protein Interactions KW - Radical SAM Enzyme KW - Recombinant Protein Expression KW - S-Adenosylmethionine (SAM) SP - 14200 EP - 11 JF - The Journal of biological chemistry JO - J. Biol. Chem. VL - 288 IS - 20 N2 - The biosynthesis of iron sulfur (FeS) clusters, their trafficking from initial assembly on scaffold proteins via carrier proteins to final incorporation into FeS apoproteins, is a highly coordinated process enabled by multiprotein systems encoded in iscRSUAhscBAfdx and sufABCDSE operons in Escherichia coli. Although these systems are believed to encode all factors required for initial cluster assembly and transfer to FeS carrier proteins, accessory factors such as monothiol glutaredoxin, GrxD, and the FeS carrier protein NfuA are located outside of these defined systems. These factors have been suggested to function both as shuttle proteins acting to transfer clusters between scaffold and carrier proteins and in the final stages of FeS protein assembly by transferring clusters to client FeS apoproteins. Here we implicate both of these factors in client protein interactions. We demonstrate specific interactions between GrxD, NfuA, and the methylthiolase MiaB, a radical S-adenosyl-L-methionine-dependent enzyme involved in the maturation of a subset of tRNAs. We show that GrxD and NfuA physically interact with MiaB with affinities compatible with an in vivo function. We furthermore demonstrate that NfuA is able to transfer its cluster in vitro to MiaB, whereas GrxD is unable to do so. The relevance of these interactions was demonstrated by linking the activity of MiaB with GrxD and NfuA in vivo. We observe a severe defect in in vivo MiaB activity in cells lacking both GrxD and NfuA, suggesting that these proteins could play complementary roles in maturation and repair of MiaB. SN - 1083-351X UR - https://www.unboundmedicine.com/medline/citation/23543739/Physical_and_functional_interactions_of_a_monothiol_glutaredoxin_and_an_iron_sulfur_cluster_carrier_protein_with_the_sulfur_donating_radical_S_adenosyl_L_methionine_enzyme_MiaB_ L2 - http://www.jbc.org/cgi/pmidlookup?view=long&pmid=23543739 DB - PRIME DP - Unbound Medicine ER -