A screen of random sequences for those that alter the trafficking of the influenza virus hemagglutinin in vivo.
In order to determine if the sequence patterns known to specify internalization represent the majority of possible internalization signals, we identified random sequences capable of causing a reporter protein to be internalized at least several-fold faster than the rate of non-selective internalization of membrane by clathrin-coated pits. A library of influenza hemagglutinin (HA) proteins, bearing short random sequences in place of the wild-type cytoplasmic domain, was prepared in recombinant SV40 virus. The library was expressed and screened for HAs that could internalize anti-HA antibody from the medium. The cytoplasmic sequences of the selected proteins were determined. From a small sample of sequences we detected several that did not resemble those previously identified. The known internalization signals must represent only a subset of the sequences that can serve as internalization signals.
Lewis
C M
CM
Department of Biochemistry, University of Texas Southwestern Medical Center at Dallas, Dallas, TX 75235-9038, USA.
Latham
K
K
Roth
M G
MG
D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D000818:Animals; D002460:Cell Line / Q000378:metabolism; D002522:Chlorocebus aethiops; D022163:Clathrin-Coated Vesicles / Q000378:metabolism; D003062:Codon / Q000235:genetics; D004705:Endocytosis / Q000502:physiology*; D011992:Endosomes / Q000378:metabolism; D005347:Fibroblasts / Q000378:metabolism; D015723:Gene Library; D017930:Genes, Reporter; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D009980:Influenza A virus / Q000235:genetics; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D017434:Protein Structure, Tertiary; D021381:Protein Transport; D011897:Random Allocation; D011993:Recombinant Fusion Proteins / Q000378:metabolism
D003062:Codon; D019267:Hemagglutinin Glycoproteins, Influenza Virus; D011993:Recombinant Fusion Proteins