Different legumin protein domains act as vacuolar targeting signals.
Legumin subunits are synthesized as precursor polypeptides and are transported into protein storage vacuoles in field bean cotyledons. We expressed a legumin subunit in yeast and found that in these cells it is also transported into the vacuoles. To elucidate vacuolar targeting information, we constructed gene fusions of different legumin propolypeptide segments with either yeast invertase or chloramphenicol acetyltransferase as reporters for analysis in yeast or plant cells, respectively. In yeast, increasing the length of the amino-terminal segment increased the portion of invertase directed to the vacuole. Only the complete legumin alpha chain (281 amino acids) directed over 90% to the vacuole. A short carboxy-terminal legumin segment (76 amino acids) fused to the carboxy terminus of invertase also efficiently targeted this fusion product to yeast vacuoles. With amino-terminal legumin-chloramphenicol acetyltransferase fusions expressed in tobacco seeds, efficient vacuolar targeting was obtained only with the complete alpha chain. We conclude that legumin contains multiple targeting information, probably formed by higher structures of relatively long peptide sequences.
Saalbach
G
G
Institute of Genetics and Crop Plant Research, Gatersleben, Sachsen-Anhalt, Federal Republic of Germany.
Jung
R
R
Kunze
G
G
Saalbach
I
I
Adler
K
K
Müntz
K
K
D000595:Amino Acid Sequence; D001692:Biological Transport; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics / Q000378:metabolism; D007887:Fabaceae / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D017343:Genes, Plant; D006026:Glycoside Hydrolases / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D010940:Plant Proteins; D014674:Plant Proteins, Dietary / Q000235:genetics / Q000378:metabolism*; D010946:Plants, Medicinal; D010947:Plants, Toxic; D011498:Protein Precursors / Q000235:genetics / Q000378:metabolism*; D021382:Protein Sorting Signals / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*; D012639:Seeds / Q000378:metabolism; D013329:Structure-Activity Relationship; D014026:Nicotiana / Q000235:genetics / Q000378:metabolism; D014617:Vacuoles / Q000378:metabolism*; D043324:beta-Fructofuranosidase; D000097793:Legumins
D015500:Chloramphenicol O-Acetyltransferase; D006026:Glycoside Hydrolases; D010940:Plant Proteins; D014674:Plant Proteins, Dietary; D011498:Protein Precursors; D021382:Protein Sorting Signals; D011994:Recombinant Proteins; D043324:beta-Fructofuranosidase; D000097793:Legumins