Literature summary for 1.3.1.B13 extracted from

Jozwiak, A.; Gutkowska, M.; Gawarecka, K.; Surmacz, L.; Buczkowska, A.; Lichocka, M.; Nowakowska, J.; Swiezewska, E.
Polyprenol reductase2 deficiency is lethal in Arabidopsis due to male sterility (2015), Plant Cell, 27, 3336-3353 .

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Cloned(Commentary)

Cloned (Comment)	Organism
recombinant expression of isozyme PPRD1 in Saccharomyces cerevisiae strains BY4741, L5366, and mutant strains dfg10DELTA and dfg10-100 from lasmid pYES-DEST52, recombinant expression of isozyme PPRD1 in Escherichia coli strain BL21, method optimization. Agrobacterium-mediated transient coexpression of GFP-tagged isozyme with plant organelle markers (mCherry fusions) in Nicotiana benthamiana. Quantitative PCR enzyme expression and promoter activity analysis	Arabidopsis thaliana
recombinant expression of isozyme PPRD1 in Saccharomyces cerevisiae strains BY4741, L5366, and mutant strains dfg10DELTA and dfg10-100 from lasmid pYES-DEST52, recombinant expression of wild-type an dmutant isozyme PPRD1s in Escherichia coli strain BL21, method optimization. Agrobacterium-mediated transient coexpression of GFP-tagged isozyme with plant organelle markers (mCherry fusions) in Nicotiana benthamiana. Quantitative PCR enzyme expression and promoter activity analysis	Arabidopsis thaliana

Cloned (Comment)

Organism

recombinant expression of isozyme PPRD1 in Saccharomyces cerevisiae strains BY4741, L5366, and mutant strains dfg10DELTA and dfg10-100 from lasmid pYES-DEST52, recombinant expression of isozyme PPRD1 in Escherichia coli strain BL21, method optimization. Agrobacterium-mediated transient coexpression of GFP-tagged isozyme with plant organelle markers (mCherry fusions) in Nicotiana benthamiana. Quantitative PCR enzyme expression and promoter activity analysis

Arabidopsis thaliana

recombinant expression of isozyme PPRD1 in Saccharomyces cerevisiae strains BY4741, L5366, and mutant strains dfg10DELTA and dfg10-100 from lasmid pYES-DEST52, recombinant expression of wild-type an dmutant isozyme PPRD1s in Escherichia coli strain BL21, method optimization. Agrobacterium-mediated transient coexpression of GFP-tagged isozyme with plant organelle markers (mCherry fusions) in Nicotiana benthamiana. Quantitative PCR enzyme expression and promoter activity analysis

Arabidopsis thaliana

Protein Variants

Protein Variants	Comment	Organism
H321L	site-directed mutagenesis, the mutant shows 47% of wild-type reductase activity	Arabidopsis thaliana
H336L	site-directed mutagenesis, the mutant shows 45% of wild-type reductase activity	Arabidopsis thaliana
additional information	PPRD2 mutants with His321 or His336, corresponding to His296 or His309 in human hSRD5A3, substituted by Leu are employed to complement the yeast dfg10DELTA mutant. Construction of two heterozygous T-DNA PPRD2 insertion lines, pprd2-1 (SALK_113221C) and pprd2-2 (SALK_006421), PPRD2 deficiency is lethal in Arabidopsis thaliana due to male sterility, PPRD2 knockout plants are not viable. For the pprd2 homozygous mutants, consistently with their Dol deficiency, defects in N-glycosylation are accompanied by degradation of some proteins, e.g., SKU5	Arabidopsis thaliana
additional information	total loss of activity of the putative C-terminally truncated PPRD1 proteins encoded by PPRD1-INT3 and PPRD1-INT4	Arabidopsis thaliana

Localization

Localization	Comment	Organism	GeneOntology No.	Textmining
endoplasmic reticulum	-	Arabidopsis thaliana	5783	-
additional information	polyprenol reductases are diversely localized within the plant cell, isozyme expression analysis, overview	Arabidopsis thaliana	-	-
plasma membrane	-	Arabidopsis thaliana	5886	-

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
a polyprenol + NADPH + H+	Arabidopsis thaliana	-	a dolichol + NADP+	-	?
a polyprenol + NADPH + H+	Arabidopsis thaliana Col-0	-	a dolichol + NADP+	-	?

Organism

Organism	UniProt	Comment	Textmining
Arabidopsis thaliana	Q9CAH5	-	-
Arabidopsis thaliana	Q9SI62	-	-
Arabidopsis thaliana Col-0	Q9CAH5	-	-
Arabidopsis thaliana Col-0	Q9SI62	-	-
no activity in Escherichia coli	-	-	-

Source Tissue

Source Tissue	Comment	Organism	Textmining
flower	-	Arabidopsis thaliana	-
flower bud	-	Arabidopsis thaliana	-
leaf	-	Arabidopsis thaliana	-
additional information	polyprenol reductases are expressed in a tissue-specific manner in Arabidopsis thaliana. Isozyme PPRD1 is expressed in young seedlings, while in older plants, it is mainly expressed in the roots and flowers. Expression of PPRD1 increases in the roots and decreases in the leaves. Isozyme expression analysis, overview	Arabidopsis thaliana	-
additional information	polyprenol reductases are expressed in a tissue-specific manner in Arabidopsis thaliana. Isozyme PRD2 is expressed in all organs analyzed in young seedlings and older plants. With age, expression of PPRD2 is fairly constant in the roots and decreases in the leaves. Isozyme expression analysis, overview	Arabidopsis thaliana	-
pollen	only isozyme PPRD2	Arabidopsis thaliana	-
root	-	Arabidopsis thaliana	-
stem	-	Arabidopsis thaliana	-

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
a polyprenol + NADPH + H+	-	Arabidopsis thaliana	a dolichol + NADP+	-	?
a polyprenol + NADPH + H+	-	Arabidopsis thaliana Col-0	a dolichol + NADP+	-	?

Synonyms

Synonyms	Comment	Organism
polyprenol reductase 1	-	Arabidopsis thaliana
polyprenol reductase 2	-	Arabidopsis thaliana
polyprenol reductase2	-	Arabidopsis thaliana
PPRD1	-	Arabidopsis thaliana
PPRD2	-	Arabidopsis thaliana

Cofactor

Cofactor	Comment	Organism	Structure
NADPH	-	Arabidopsis thaliana

General Information

General Information	Comment	Organism
evolution	the two genes PPRD1 and PPRD2 are orthologous to human SRD5A3 (steroid 5alpha reductase type 3) and encode polyprenol reductases responsible for conversion of polyprenol to dolichol in Arabidopsis thaliana	Arabidopsis thaliana
malfunction	polyprenol reductase2 deficiency is lethal in Arabidopsis thaliana due to male sterility, PPRD2 knockout plants are not viable. PPRD2 deficiency results in aberrant development of the male gametophyte and sporophyte. Impaired protein glycosylation seems to be the major factor underlying these defects although disturbances in other cellular dolichol-dependent processes might also contribute. Shortage of dolichol in PPRD2-deficient cells is partially rescued by PPRD1 overexpression or by supplementation with dolichol. Supplementation of the human diet with dolichol-enriched plant tissues can allow therapeutic interventions in glycosylation disorders	Arabidopsis thaliana
additional information	the catalytic 3-oxo-5alpha-steroid 4-dehydrogenase domain of PPRD is localized at the C-terminus	Arabidopsis thaliana
additional information	the catalytic 3-oxo-5alpha-steroid domain of PPRD is localized at the C-terminus. Besides His321 or His336, other residues of PPRD2 might be involved in the reduction	Arabidopsis thaliana
physiological function	isozyme PPRD2 is essential for plant viability	Arabidopsis thaliana
physiological function	isozymes PPRD1 and PPRD2 are the factors mediating the key step of the dolichol cycle in plant cells. Shortage of dolichol in PPRD2-deficient cells is partially rescued by PPRD1 overexpression	Arabidopsis thaliana