We found a match
Your institution may have rights to this item. Sign in to continue.
- Title
Bioinformatic and experimental evidence for suicidal and catalytic plant THI4s.
- Authors
Joshi, Jaya; Beaudoin, Guillaume A. W.; Patterson, Jenelle A.; García-García, Jorge D.; Belisle, Catherine E.; Lan-Yen Chang; Lei Li; Duncan, Owen; Millar, A. Harvey; Hanson, Andrew D.
- Abstract
Like fungi and some prokaryotes, plants use a thiazole synthase (THI4) to make the thiazole precursor of thiamin. Fungal THI4s are suicide enzymes that destroy an essential active-site Cys residue to obtain the sulfur atom needed for thiazole formation. In contrast, certain prokaryotic THI4s have no active-site Cys, use sulfide as sulfur donor, and are truly catalytic. The presence of a conserved active-site Cys in plant THI4s and other indirect evidence implies that they are suicidal. To confirm this, we complemented the Arabidopsis tz-1 mutant, which lacks THI4 activity, with a His-tagged Arabidopsis THI4 construct. LC-MS analysis of tryptic peptides of the THI4 extracted from leaves showed that the active-site Cys was predominantly in desulfurated form, consistent with THI4 having a suicide mechanism in planta. Unexpectedly, transcriptome data mining and deep proteome profiling showed that barley, wheat, and oat have both a widely expressed canonical THI4 with an active-site Cys, and a THI4-like paralog (non-Cys THI4) that has no active-site Cys and is the major type of THI4 in developing grains. Transcriptomic evidence also indicated that barley, wheat, and oat grains synthesize thiamin de novo, implying that their non-Cys THI4s synthesize thiazole. Structure modeling supported this inference, as did demonstration that non-Cys THI4s have significant capacity to complement thiazole auxotrophy in Escherichia coli. There is thus a prima facie case that non-Cys cereal THI4s, like their prokaryotic counterparts, are catalytic thiazole synthases. Bioenergetic calculations show that, relative to suicide THI4s, such enzymes could save substantial energy during the grain-filling period.
- Subjects
OATS; PLANTS; SYNTHASES; BARLEY; DATA mining; WHEAT; SUICIDE statistics
- Publication
Biochemical Journal, 2020, Vol 477, Issue 11, p2055
- ISSN
0264-6021
- Publication type
Article
- DOI
10.1042/BCJ20200297