Literature summary for 1.14.19.35 extracted from

Venegas-Caleron, M.; Beaudoin, F.; Garces, R.; Napier, J.A.; Martinez-Force, E.
The sunflower plastidial omega3-fatty acid desaturase (HaFAD7) contains the signalling determinants required for targeting to, and retention in, the endoplasmic reticulum membrane in yeast but requires co-expressed ferredoxin for activity (2010), Phytochemistry, 71, 1050-1058.

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Cloned(Commentary)

Cloned (Comment)	Organism
gene fad7, sequence comparison with FAD3, recombinant expression in Saccharomyces cerevisiae strain W303-1A, plastidial omega3-fatty acid desaturase contains the signalling determinants required for targeting to, and retention in, the endoplasmic reticulum membrane in Saccharomyces cerevisiae but requires co-expressed ferredoxin for activity. Heterologous enzyme expression in yeast shows low activity in contrast to similar expression of microsomal FAD3 omega 3-desaturases. Expression of enzyme variants N-terminally tagged with HA, and/or anchor domains ELO or FAE, and containing a KKNL motif at the C-terminus. Deletion of the endogenous KEK C-terminal motif results in dramatically reduced accumulation of FAD7-HA. Coexpression of ferredoxin	Helianthus annuus

Cloned (Comment)

Organism

gene fad7, sequence comparison with FAD3, recombinant expression in Saccharomyces cerevisiae strain W303-1A, plastidial omega3-fatty acid desaturase contains the signalling determinants required for targeting to, and retention in, the endoplasmic reticulum membrane in Saccharomyces cerevisiae but requires co-expressed ferredoxin for activity. Heterologous enzyme expression in yeast shows low activity in contrast to similar expression of microsomal FAD3 omega 3-desaturases. Expression of enzyme variants N-terminally tagged with HA, and/or anchor domains ELO or FAE, and containing a KKNL motif at the C-terminus. Deletion of the endogenous KEK C-terminal motif results in dramatically reduced accumulation of FAD7-HA. Coexpression of ferredoxin

Helianthus annuus

Protein Variants

Protein Variants	Comment	Organism
additional information	the removal of the plastidial transit peptide and the incorporation of a KKNL motif to the C-terminus of HaFAD7 increases the activity by 10fold compared to the native protein. N-terminal fusion of transmembrane-domains from either the Saccharomyces cerevisiae microsomal ELO3, (a type III signal anchor domain), or FAE1, an endoplasmic reticulum membrane anchoring domain, results in moderate increases in enzyme activity. Fusing a hemagglutinin epitope tag upstream of an endogenous C-terminal KEK motif results in a significant loss of activity compared to the un-tagged construct, indicating that the endogenous KEK C-terminal di-lysine motif is capable of directing in yeast the ER-retention of this normally plastidial-located protein	Helianthus annuus

Localization

Localization	Comment	Organism	GeneOntology No.	Textmining
chloroplast membrane	-	Helianthus annuus	31969	-

Organism

Organism	UniProt	Comment	Textmining
Helianthus annuus	Q56VS4	gene fad7	-

Source Tissue

Source Tissue	Comment	Organism	Textmining

Synonyms

Synonyms	Comment	Organism
FAD7 omega 3-desaturase	-	Helianthus annuus
HaFAD7	-	Helianthus annuus

Cofactor

Cofactor	Comment	Organism	Structure
Ferredoxin	required for activity in the chloroplasts, ferredoxin is required for recombinantly expressed HaFAD7 activity but not for protein stability and accumulation in yeast	Helianthus annuus