Peroxisomal amine oxidase of Hansenula polymorpha does not require its SRL-containing C-terminal sequence for targeting.
Amine oxidase (AMO) is a peroxisomal matrix protein of Hansenula polymorpha, which is induced during growth of the yeast in media containing primary amines as a sole nitrogen source. The deduced amino acid sequence of the protein contains an SRL sequence at nine amino acids from the C-terminus. In this study, we have examined the possible role of the SRL motif in sorting of AMO to peroxisomes by mutating the corresponding gene sequence. For this purpose, we have developed a DNA construct that is specifically integrated into the AMO locus of the H. polymorpha genome, placing the mutant gene under the control of the endogenous AMO promoter and eliminating expression of the wild-type gene. Analysis of a stable transformant, containing the desired gene configuration, showed that mutation of the C-terminal sequence neither interfered with correct targeting of the protein into the peroxisome nor displayed significant effects on its activity. From this, it was concluded that the SRL-containing C-terminus is not essential for peroxisomal targeting of AMO in H. polymorpha.
Faber
K N
KN
Department of Biochemistry, Groningen University, The Netherlands.
Haima
P
P
de Hoop
M J
MJ
Harder
W
W
Veenhuis
M
M
Ab
G
G
D006631:Amine Oxidase (Copper-Containing); D000595:Amino Acid Sequence; D001483:Base Sequence; D001692:Biological Transport; D002451:Cell Compartmentation; D006651:Histocytochemistry; D007150:Immunohistochemistry; D008830:Microbodies / Q000201:enzymology* / Q000648:ultrastructure; D008969:Molecular Sequence Data; D016296:Mutagenesis; D000587:Oxidoreductases Acting on CH-NH Group Donors / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D010843:Pichia / Q000201:enzymology* / Q000235:genetics / Q000648:ultrastructure; D021382:Protein Sorting Signals / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism; D014170:Transformation, Genetic
D021382:Protein Sorting Signals; D011994:Recombinant Proteins; D006631:Amine Oxidase (Copper-Containing); D000587:Oxidoreductases Acting on CH-NH Group Donors