Role of a conserved amino-terminal sequence in the ecotropic MLV receptor mCAT1.
The amino-terminus of mCAT1 and homologous proteins is predicted to form a positively charged, amphipathic alpha helix on the cytoplasmic side of the plasma membrane. Peptides with similar sequence motifs often provide membrane anchors, protein-protein interaction domains, or intracellular transport-targeting signals. Deleting most of the cytoplasmic N-terminal sequence of mCAT1 led to reduced expression on the cell surface and accumulation in the endoplasmic reticulum but did not abrogate receptor function. Surprisingly, when the N-terminal 36 or 18 amino acids of mCAT1 were fused to green fluorescent protein (gfp), gfp accumulated almost exclusively in mitochondria. Mitochondrial targeting depended on arginines at positions 15 and 16 and was inhibitable by downstream transmembrane sequences. Although the full-length mCAT1 was not detected in mitochondria, the mitochondrial-targeting property of the N-terminal sequence fused to gfp is conserved in orthologous and paralogous proteins that diverged approximately 80 million years ago, suggesting a conserved biological function. We propose that the conserved N-terminal motif of CAT proteins provides a regulatable signal for transport to, or retention in, different cell membrane compartments.
Ou
Wu
W
Laboratory of Molecular Microbiology, National Institute of Allergy and Infectious Disease, National Institutes of Health, Bethesda, MD 20892, USA.
Silver
Jonathan
J
D020816:Amino Acid Motifs / Q000235:genetics / Q000502:physiology*; D000818:Animals; D001120:Arginine; D027201:Cationic Amino Acid Transporter 1 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D006224:Cricetinae; D004721:Endoplasmic Reticulum / Q000378:metabolism; D017353:Gene Deletion; D049452:Green Fluorescent Proteins; D009052:Leukemia Virus, Murine / Q000502:physiology*; D008164:Luminescent Proteins / Q000737:chemistry; D008562:Membrane Glycoproteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008928:Mitochondria / Q000378:metabolism; D021382:Protein Sorting Signals; D021381:Protein Transport; D011991:Receptors, Virus / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011995:Recombination, Genetic; D020079:Terminal Repeat Sequences / Q000502:physiology*
D027201:Cationic Amino Acid Transporter 1; D008164:Luminescent Proteins; D008562:Membrane Glycoproteins; D021382:Protein Sorting Signals; D011991:Receptors, Virus; C420826:Slc7a1 protein, mouse; C070506:ecotropic murine leukemia virus receptor; D049452:Green Fluorescent Proteins; D001120:Arginine