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Category:MichSt14A 1

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StatusPageUserDate/TimeGO Term (Aspect)ReferenceEvidenceNotesLinks
acceptableCHLRE:A1YSB4Amstutzc, MichSt14A 12014-04-04 10:14:46 CDTGO:0009579 thylakoid (C)PMID:19574473ECO:0000314 direct assay evidence used in manual assertion

In Figure 5, wild type, rep27, and rep27 complimented cells were fractionated into soluble and membrane portions and analyzed for the presence of REP27. The western blot analysis detected REP27 in the total membrane and total extract fractions for the wild type and complemented cells. However no detection was found in the soluble fraction. Figure 6 isolated thylakoid memebranes and again looked for the presence of REP27 in the membrane and soluble fractions. REP27 was present in all membrane fractions but was absent from the soluble. These two figures indicate that the REP27 protein is localized to the thylakoid membranes in the chloroplast.

requireschanges?Amstutzc, MichSt14A 12014-04-03 20:53:19 CDTGO:0005634 nucleus (In supplemental figure 4, a GUS reporter was tagged to PIF1. The same cells were also stained with a DNA stain, DAPI, which localizes the nucleus. When the two images were superimposed, the GUS-PIF1 was localized only to the region that was also stained with DAPI, indicating PIF1 is in the nucleus.)PMID:15448264IDA: Inferred from Direct Assay
requireschangesARATH:ZEPAmstutzc, MichSt14A 12014-04-03 13:04:01 CDTGO:0010028 xanthophyll cycle (P)PMID:12237386ECO:0000315 mutant phenotype evidence used in manual assertion

The assignment of this protein to an activity is an infererence from the mutant phenotype related to accumulation of different compounds "the Npq- phenotype of npq2 co- segregated with the accumulation of zeaxanthin and absence of antheraxanthin, violaxanthin, and neoxanthin (data not shown)." According to figure 6, the xanthophyll pool in wild type cells in low light is primarily violaxanthin. In high light this violaxanthin decreases, producing antheraxanthin and zeaxanthin. However, the amount of zeaxanthin in the NPQ2 mutant remains the same when under low light. There is no production of antheraxanin or violaxanthin. This is due to the lack of zeaxanthin epoxidase activity.

requireschanges?Amstutzc, MichSt14A 12014-04-03 12:45:24 CDTGO:0046422 violaxanthin de-epoxidase activity (Figure 6 shows that under low light the pool of xanthophyll pigments in wild type Chlamydomonas reinhardtii is primarily composed of violxanthin. Under high light conditions, violaxanthin is epoxidated to antheraxanthin and zeaxanthin. However in the mutant NPQ1, the xanthin pool under high light remains primarily violaxanthin. Therefore the mutant is lacking violaxanthin epoxidase activity.)PMID:12237386IMP: Inferred from Mutant Phenotype
acceptableCHLRE:STT7Amstutzc, MichSt14A 12014-04-03 11:07:49 CDTGO:0004674 protein serine/threonine kinase activity (F)PMID:12624266ECO:0000315 mutant phenotype evidence used in manual assertion

Figure 1 shows that stt7 Chlamydomonas reinhardtii mutants have increased maximal fluorescence compared to wild type under conditions that would induce state II transition. This mutant phenotype is rescued when complimented with the Stt7 gene. Thylakoid protein fractions with radiolabeled phosphate under state I and state II transition show that the phosphorylation status increases in transition II in wild type and rescued cells, but not in the stt7 mutants. Western blotting of similar fractions reacted with phosphor-threonine antiserum showed similar results. Since the antiserum will only bind to phosphate groups on threonine residues, the protein must be a threonine/serine kinase.


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