Chloroplast iron-sulfur cluster protein matu.pdf

Chloroplast iron-sulfur cluster protein maturation requires the essential cysteine desulfurase CpNifS Douglas Van Hoewyk*, Salah E. Abdel-Ghany*, Christopher M. Cohu*, Stephen K. Herbert?, Paul Kugrens*, Marinus Pilon*, and Elizabeth A. H. Pilon-Smits*? *Biology Department and Program in Molecular Plant Biology, Colorado State University, Fort Collins, CO, 80526-1878; and ?Department of Botany, University of Wyoming, 1000 East University Avenue, Laramie, WY 82071 Edited by Bob B. Buchanan, University of California, Berkeley, CA, and approved February 12, 2007 (received for review January 27, 2007) NifS-like proteins provide the sulfur (S) for the formation of iron-sulfur (Fe-S) clusters, an ancient and essential type of cofactor found in all three domains of life. Plants are known to contain two distinct NifS-like proteins, localized in the mitochondria (MtNifS) and the chloroplast (CpNifS). In the chloroplast, five different Fe-S cluster types are required in various proteins. These plastid Fe-S proteins are involved in a variety of biochemical pathways includ- ing photosynthetic electron transport and nitrogen and sulfur assimilation. In vitro, the chloroplastic cysteine desulfurase CpNifS can release elemental sulfur from cysteine for Fe-S cluster biogen- esis in ferredoxin. However, because of the lack of a suitable mutant allele, the role of CpNifS has not been studied thus far in planta. To study the role of CpNifS in Fe-S cluster biogenesis in vivo, the gene was silenced by using an inducible RNAi (interference) approach. Plants with reduced CpNifS expression exhibited chlo- rosis, a disorganized chloroplast structure, and stunted growth and eventually became necrotic and died before seed set. Photosyn- thetic electron transport and carbon dioxide assimilation were severely impaired in the silenced plant lines. The silencing of CpNifS decreased the abundance of all chloroplastic Fe-S proteins tested, representing all five Fe-S cluster types. Mitochondrial Fe-