DNA binding and partial nucleoid localization of the chloroplast stromal enzyme ferredoxin:sulfite reductase.
Sulfite reductase (SiR) is an important enzyme catalyzing the reduction of sulfite to sulfide during sulfur assimilation in plants. This enzyme is localized in plastids, including chloroplasts, and uses ferredoxin as an electron donor. Ferredoxin-dependent SiR has been found in isolated chloroplast nucleoids, but its localization in vivo or in intact plastids has not been examined. Here, we report the DNA-binding properties of SiRs from pea (PsSiR) and maize (ZmSiR) using an enzymatically active holoenzyme with prosthetic groups. PsSiR binds to both double-stranded and single-stranded DNA without significant sequence specificity. DNA binding did not affect the enzymatic activity of PsSiR, suggesting that ferredoxin and sulfite are accessible to SiR molecules within the nucleoids. Comparison of PsSiR and ZmSiR suggests that ZmSiR does indeed have DNA-binding activity, as was reported previously, but the DNA affinity and DNA-compacting ability are higher in PsSiR than in ZmSiR. The tight compaction of nucleoids by PsSiR led to severe repression of transcription activity in pea nucleoids. Indirect immunofluorescence microscopy showed that the majority of SiR molecules colocalized with nucleoids in pea chloroplasts, whereas no particular localization to nucleoids was detected in maize chloroplasts. These results suggest that SiR plays an essential role in compacting nucleoids in plastids, but that the extent of association of SiR with nucleoids varies among plant species.
Sekine
Kohsuke
K
Department of Molecular Biology, Faculty of Science, Saitama University, Japan.
Fujiwara
Makoto
M
Nakayama
Masato
M
Takao
Toshifumi
T
Hase
Toshiharu
T
Sato
Naoki
N
D000595:Amino Acid Sequence; D022003:Cell Nucleus Structures / Q000201:enzymology; D002736:Chloroplasts / Q000201:enzymology* / Q000648:ultrastructure; D004247:DNA / Q000378:metabolism*; D008969:Molecular Sequence Data; D018532:Pisum sativum / Q000201:enzymology; D050889:Sulfite Reductase (Ferredoxin) / Q000032:analysis* / Q000737:chemistry / Q000235:genetics; D014544:Uridine Triphosphate / Q000378:metabolism; D003313:Zea mays / Q000201:enzymology
D004247:DNA; D050889:Sulfite Reductase (Ferredoxin); D014544:Uridine Triphosphate