HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Palenchar, P. M. Articles by Mueller, E. G. Search for Related Content PubMed PubMed Citation Articles by Palenchar, P. M. Articles by Mueller, E. G. Social Bookmarking                      What's this?

J Biol Chem, Vol. 275, Issue 12, 8283-8286, March 24, 2000

ACCELERATED PUBLICATION
Evidence That ThiI, an Enzyme Shared between Thiamin and 4-Thiouridine Biosynthesis, May Be a Sulfurtransferase That Proceeds through a Persulfide Intermediate^*

Peter M. Palenchar^§, Christopher J. Buck, Hui Cheng^¶, Timothy J. Larson^¶, and Eugene G. Mueller

From the  Department of Chemistry and Biochemistry, University of Delaware, Newark, Delaware 19716 and the ^¶ Department of Biochemistry, Virginia Polytechnic Institute and State University, Blacksburg, Virginia 24061

ThiI is an enzyme common to the biosynthetic pathways leading to both thiamin and 4-thiouridine in tRNA. Comparison of the ThiI sequence with protein sequences in the data bases revealed that the Escherichia coli enzyme contains a C-terminal extension displaying sequence similarity to the sulfurtransferase rhodanese. Cys-456 of ThiI aligns with the active site cysteine residue of rhodanese that transiently forms a persulfide during catalysis. We investigated the functional importance of this sequence similarity and discovered that, like rhodanese, ThiI catalyzes the transfer of sulfur from thiosulfate to cyanide. Mutation of Cys-456 to alanine impairs this sulfurtransferase activity, and the C456A ThiI is incapable of supporting generation of 4-thiouridine in tRNA both in vitro and in vivo. We therefore conclude that Cys-456 of ThiI is critical for activity and propose that Cys-456 transiently forms a persulfide during catalysis. To accommodate this hypothesis, we propose a general mechanism for sulfur transfer in which the terminal sulfur of the persulfide first acts as a nucleophile and is then transferred as an equivalent of S² rather than S⁰.

---

^* This work was supported in part by the National Institutes of Health Grant GM59636-01 (to E. G. M.) and National Science Foundation Grant MCB-9118757 (to T. J. L.).The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

^§ Supported in part by United States Public Health Service Grant T32 GM08550.

To whom correspondence should be addressed: Dept. of Chemistry & Biochemistry, University of Delaware, Newark, DE 19716. Tel.: 302-831-2739; Fax: 302-831-6335; E-mail: emueller@udel.edu.

---

Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.

CiteULike

Complore

Connotea

Del.icio.us

Digg

Reddit

Technorati

This article has been cited by other articles:

				B. Campanini, F. Schiaretti, S. Abbruzzetti, D. Kessler, and A. Mozzarelli Sulfur Mobilization in Cyanobacteria: THE CATALYTIC MECHANISM OF L-CYSTINE C-S LYASE (C-DES) FROM SYNECHOCYSTIS J. Biol. Chem., December 15, 2006; 281(50): 38769 - 38780. [Abstract] [Full Text] [PDF]

				N. Shigi, T. Suzuki, T. Terada, M. Shirouzu, S. Yokoyama, and K. Watanabe Temperature-dependent Biosynthesis of 2-Thioribothymidine of Thermus thermophilus tRNA J. Biol. Chem., January 27, 2006; 281(4): 2104 - 2113. [Abstract] [Full Text] [PDF]

				G. Gabant, S. Auxilien, I. Tuszynska, M. Locard, M. J. Gajda, G. Chaussinand, B. Fernandez, A. Dedieu, H. Grosjean, B. Golinelli-Pimpaneau, et al. THUMP from archaeal tRNA:m22G10 methyltransferase, a genuine autonomously folding domain. Nucleic Acids Res., January 1, 2006; 34(9): 2483 - 2494. [Abstract] [Full Text] [PDF]

				N. C. Martinez-Gomez, M. Robers, and D. M. Downs Mutational Analysis of ThiH, a Member of the Radical S-Adenosylmethionine (AdoMet) Protein Superfamily J. Biol. Chem., September 24, 2004; 279(39): 40505 - 40510. [Abstract] [Full Text] [PDF]

				J. Armengaud, J. Urbonavicius, B. Fernandez, G. Chaussinand, J. M. Bujnicki, and H. Grosjean N2-Methylation of Guanosine at Position 10 in tRNA Is Catalyzed by a THUMP Domain-containing, S-Adenosylmethionine-dependent Methyltransferase, Conserved in Archaea and Eukaryota J. Biol. Chem., August 27, 2004; 279(35): 37142 - 37152. [Abstract] [Full Text] [PDF]

				C. T. Lauhon, W. M. Erwin, and G. N. Ton Substrate Specificity for 4-Thiouridine Modification in Escherichia coli J. Biol. Chem., May 28, 2004; 279(22): 23022 - 23029. [Abstract] [Full Text] [PDF]

				C. T. Lauhon, E. Skovran, H. D. Urbina, D. M. Downs, and L. E. Vickery Substitutions in an Active Site Loop of Escherichia coli IscS Result in Specific Defects in Fe-S Cluster and Thionucleoside Biosynthesis in Vivo J. Biol. Chem., May 7, 2004; 279(19): 19551 - 19558. [Abstract] [Full Text] [PDF]

				R. Leonardi and P. L. Roach Thiamine Biosynthesis in Escherichia coli: IN VITRO RECONSTITUTION OF THE THIAZOLE SYNTHASE ACTIVITY J. Biol. Chem., April 23, 2004; 279(17): 17054 - 17062. [Abstract] [Full Text] [PDF]

				G. Jager, R. Leipuviene, M. G. Pollard, Q. Qian, and G. R. Bjork The Conserved Cys-X1-X2-Cys Motif Present in the TtcA Protein Is Required for the Thiolation of Cytidine in Position 32 of tRNA from Salmonella enterica serovar Typhimurium J. Bacteriol., February 1, 2004; 186(3): 750 - 757. [Abstract] [Full Text] [PDF]

				R. Leipuviene, Q. Qian, and G. R. Bjork Formation of Thiolated Nucleosides Present in tRNA from Salmonella enterica serovar Typhimurium Occurs in Two Principally Distinct Pathways J. Bacteriol., February 1, 2004; 186(3): 758 - 766. [Abstract] [Full Text] [PDF]

				M. D. Wolfe, F. Ahmed, G. M. Lacourciere, C. T. Lauhon, T. C. Stadtman, and T. J. Larson Functional Diversity of the Rhodanese Homology Domain: THE ESCHERICHIA COLI ybbB GENE ENCODES A SELENOPHOSPHATE-DEPENDENT tRNA 2-SELENOURIDINE SYNTHASE J. Biol. Chem., January 16, 2004; 279(3): 1801 - 1809. [Abstract] [Full Text] [PDF]

				M. S. Alphey, R. A. M. Williams, J. C. Mottram, G. H. Coombs, and W. N. Hunter The Crystal Structure of Leishmania major 3-Mercaptopyruvate Sulfurtransferase: A THREE-DOMAIN ARCHITECTURE WITH A SERINE PROTEASE-LIKE TRIAD AT THE ACTIVE SITE J. Biol. Chem., November 28, 2003; 278(48): 48219 - 48227. [Abstract] [Full Text] [PDF]

				F. Forlani, A. Carpen, and S. Pagani Evidence that elongation of the catalytic loop of the Azotobacter vinelandii rhodanese changed selectivity from sulfur- to phosphate-containing substrates Protein Eng. Des. Sel., July 1, 2003; 16(7): 515 - 519. [Abstract] [Full Text] [PDF]

				R. A. M. Williams, S. M. Kelly, J. C. Mottram, and G. H. Coombs 3-Mercaptopyruvate Sulfurtransferase of Leishmania Contains an Unusual C-terminal Extension and Is Involved in Thioredoxin and Antioxidant Metabolism J. Biol. Chem., January 10, 2003; 278(3): 1480 - 1486. [Abstract] [Full Text] [PDF]

				K. Nilsson, H. K. Lundgren, T. G. Hagervall, and G. R. Bjork The Cysteine Desulfurase IscS Is Required for Synthesis of All Five Thiolated Nucleosides Present in tRNA from Salmonella enterica Serovar Typhimurium J. Bacteriol., December 15, 2002; 184(24): 6830 - 6835. [Abstract] [Full Text] [PDF]

				J. Xi, Y. Ge, C. Kinsland, F. W. McLafferty, and T. P. Begley Biosynthesis of the thiazole moiety of thiamin in Escherichia coli: Identification of an acyldisulfide-linked protein-protein conjugate that is functionally analogous to the ubiquitin/E1 complex PNAS, June 28, 2001; (2001) 141226698. [Abstract] [Full Text] [PDF]

				C. T. Lauhon and R. Kambampati The iscS Gene in Escherichia coli Is Required for the Biosynthesis of 4-Thiouridine, Thiamin, and NAD J. Biol. Chem., June 23, 2000; 275(26): 20096 - 20103. [Abstract] [Full Text] [PDF]

				E. G. Mueller, P. M. Palenchar, and C. J. Buck The Role of the Cysteine Residues of ThiI in the Generation of 4-Thiouridine in tRNA J. Biol. Chem., August 31, 2001; 276(36): 33588 - 33595. [Abstract] [Full Text] [PDF]

				H. D. Urbina, J. J. Silberg, K. G. Hoff, and L. E. Vickery Transfer of Sulfur from IscS to IscU during Fe/S Cluster Assembly J. Biol. Chem., November 21, 2001; 276(48): 44521 - 44526. [Abstract] [Full Text] [PDF]

				J. Xi, Y. Ge, C. Kinsland, F. W. McLafferty, and T. P. Begley Biosynthesis of the thiazole moiety of thiamin in Escherichia coli: Identification of an acyldisulfide-linked protein-protein conjugate that is functionally analogous to the ubiquitin/E1 complex PNAS, July 17, 2001; 98(15): 8513 - 8518. [Abstract] [Full Text] [PDF]