Several protein regions contribute to determine the nuclear and cytoplasmic localization of the influenza A virus nucleoprotein.
A systematic analysis was carried out to identify the amino acid signals that regulate the nucleo-cytoplasmic transport of the influenza A virus nucleoprotein (NP). The analysis involved determining the intracellular localization of eight deleted recombinant NP proteins and 14 chimeric proteins containing the green fluorescent protein fused to different NP fragments. In addition, the subcellular distribution of NP derivatives that contained specific substitutions at serine-3, which is the major phosphorylation site of the A/Victoria/3/75 NP, were analysed. From the results obtained, it is concluded that the NP contains three signals involved in nuclear accumulation and two regions that cause cytoplasmic accumulation of the fusion proteins. One of the karyophilic signals was located at the N terminus of the protein, and the data obtained suggest that the functionality of this signal can be modified by phosphorylation at serine-3. These findings are discussed in the context of the transport of influenza virus ribonucleoprotein complexes into and out of the nucleus.
Bullido
R
R
Centro Nacional de Biología Fundamental, Instituto de Salud Carlos III, Majadahonda 28220, Madrid, Spain.
Gómez-Puertas
P
P
Albo
C
C
Portela
A
A
D019943:Amino Acid Substitution; D000818:Animals; D001692:Biological Transport; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism* / Q000821:virology; D003593:Cytoplasm / Q000378:metabolism* / Q000821:virology; D019084:Fluorescent Antibody Technique, Indirect; D049452:Green Fluorescent Proteins; D009980:Influenza A virus / Q000235:genetics / Q000378:metabolism*; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D019590:Nucleocapsid Proteins; D009698:Nucleoproteins; D010957:Plasmids / Q000235:genetics; D021382:Protein Sorting Signals / Q000378:metabolism*; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction; D014758:Viral Core Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
D008164:Luminescent Proteins; D019590:Nucleocapsid Proteins; D009698:Nucleoproteins; D021382:Protein Sorting Signals; D011993:Recombinant Fusion Proteins; D014758:Viral Core Proteins; D049452:Green Fluorescent Proteins