Signal-sequence trap in mammalian and yeast cells

TY - JOUR

T1 - Signal-sequence trap in mammalian and yeast cells

T2 - a comparison

AU - Galliciotti, G

AU - Schneider, H

AU - Wyder, L

AU - Vitaliti, A

AU - Wittmer, M

AU - Ajmo, M

AU - Klemenz, R

PY - 2001/10/1

Y1 - 2001/10/1

N2 - Membrane associated and secreted proteins are translated as precursors containing a signal peptide that allows protein-insertion into the membrane of the endoplasmic reticulum and is co-translationally removed in the lumen. The ability of the signal peptide to direct a polypeptide into the secretory pathway is exploited in methods developed to select cDNAs encoding such proteins. Different strategies are known in which cDNA libraries can be screened for signal peptides by the ability of the latter to rescue the translocation of signal sequence-less proteins. In one method, a cDNA library is tested for interleukin 2 receptor alpha chain translocation to the membrane in COS cells, in another one for invertase secretion from yeast. In this work, we compared the two systems by testing six mouse signal peptides in COS and yeast cells. All of them were functional in the mammalian system, whereas only three of them in yeast. Two other sequences needed the 5' cDNA sequence flanking the ATG codon to be removed in order to enable protein translocation. Although the structure of signal sequences and the functioning of the secretory machinery are well conserved from prokaryotes to eukaryotes, it seems evident that not all signal peptides can be interchanged between different proteins and organisms. In particular, signal peptides that are functional in the mammalian system do not necessarily lead to protein translocation in yeast.

AB - Membrane associated and secreted proteins are translated as precursors containing a signal peptide that allows protein-insertion into the membrane of the endoplasmic reticulum and is co-translationally removed in the lumen. The ability of the signal peptide to direct a polypeptide into the secretory pathway is exploited in methods developed to select cDNAs encoding such proteins. Different strategies are known in which cDNA libraries can be screened for signal peptides by the ability of the latter to rescue the translocation of signal sequence-less proteins. In one method, a cDNA library is tested for interleukin 2 receptor alpha chain translocation to the membrane in COS cells, in another one for invertase secretion from yeast. In this work, we compared the two systems by testing six mouse signal peptides in COS and yeast cells. All of them were functional in the mammalian system, whereas only three of them in yeast. Two other sequences needed the 5' cDNA sequence flanking the ATG codon to be removed in order to enable protein translocation. Although the structure of signal sequences and the functioning of the secretory machinery are well conserved from prokaryotes to eukaryotes, it seems evident that not all signal peptides can be interchanged between different proteins and organisms. In particular, signal peptides that are functional in the mammalian system do not necessarily lead to protein translocation in yeast.

KW - Animals

KW - Base Sequence

KW - COS Cells

KW - Endothelium, Vascular

KW - Eukaryotic Cells

KW - Gene Library

KW - Glycoside Hydrolases

KW - Mammals

KW - Membrane Proteins

KW - Mice

KW - Molecular Sequence Data

KW - Peptides

KW - Protein Sorting Signals

KW - Protein Transport

KW - Receptors, Interleukin-2

KW - Species Specificity

KW - Yeasts

KW - beta-Fructofuranosidase

M3 - SCORING: Journal article

C2 - 11696859

VL - 183

SP - 175

EP - 182

JO - J MEMBRANE BIOL

JF - J MEMBRANE BIOL

SN - 0022-2631

IS - 3

ER -