Skip to main content
NCBI home page
Journal of Virology logoLink to Journal of Virology
. 1984 Jan;49(1):223–228. doi: 10.1128/jvi.49.1.223-228.1984

Influenza virus hemagglutinin containing an altered hydrophobic carboxy terminus accumulates intracellularly.

M M Sveda, L J Markoff, C J Lai
PMCID: PMC255445  PMID: 6690711

Abstract

The influenza virus hemagglutinin (HA) glycoprotein synthesized from cloned DNA in a simian virus 40 vector is expressed on the surface of infected primate cells. Previously, it has been demonstrated that mutant HAs lacking the hydrophobic carboxy terminus fail to anchor on the cell surface and therefore are secreted extracellularly. During analysis of additional HA deletion mutants derived from an HA-simian virus 40 recombinant, we found a mutant with an altered hydrophobic carboxy terminus that exhibited another phenotype. This deletion mutant, dl-12, produced HA that was neither secreted nor expressed on the infected cell surface. The mutant HA was similar to the wild-type HA in apparent molecular weight and extent of glycosylation as assayed by endoglycosidase H sensitivity. The mutant HA localized near the perinuclear region of infected cells as indicated by an indirect immunofluorescence assay. Sequence analysis showed that a 5-base-pair deletion had occurred before the region encoding the hydrophobic carboxy terminus. Nevertheless, the physicochemical properties of the wild-type HA carboxy terminus were maintained in that the truncated HA carboxy terminus consisted of predominantly hydrophobic amino acids followed by several charged amino acids residues. This similarity in the carboxy terminus between the wild-type and mutant HAs may be responsible for the functional similarities observed. In spite of these similarities, the mutant HA failed to mature at the surface. These results suggest that the maturation of the mutant HA is blocked during a late stage in the transit to the cell surface.

Full text

PDF
223

Images in this article

224Image
on p.224
225Image
on p.225
225Image
on p.225
226Image
on p.226

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Blobel G., Dobberstein B. Transfer of proteins across membranes. I. Presence of proteolytically processed and unprocessed nascent immunoglobulin light chains on membrane-bound ribosomes of murine myeloma. J Cell Biol. 1975 Dec;67(3):835–851. doi: 10.1083/jcb.67.3.835. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Compans R. W. Distinct carbohydrate components of influenza virus glycoproteins in smooth and rough cytoplasmic membranes. Virology. 1973 Oct;55(2):541–545. doi: 10.1016/0042-6822(73)90199-2. [DOI] [PubMed] [Google Scholar]
  3. Compans R. W. Influenza virus proteins. II. Association with components of the cytoplasm. Virology. 1973 Jan;51(1):56–70. doi: 10.1016/0042-6822(73)90365-6. [DOI] [PubMed] [Google Scholar]
  4. Fields S., Winter G., Brownlee G. G. Structure of the neuraminidase gene in human influenza virus A/PR/8/34. Nature. 1981 Mar 19;290(5803):213–217. doi: 10.1038/290213a0. [DOI] [PubMed] [Google Scholar]
  5. Gething M. J., Bye J., Skehel J., Waterfield M. Cloning and DNA sequence of double-stranded copies of haemagglutinin genes from H2 and H3 strains elucidates antigenic shift and drift in human influenza virus. Nature. 1980 Sep 25;287(5780):301–306. doi: 10.1038/287301a0. [DOI] [PubMed] [Google Scholar]
  6. Hirst G. K. THE QUANTITATIVE DETERMINATION OF INFLUENZA VIRUS AND ANTIBODIES BY MEANS OF RED CELL AGGLUTINATION. J Exp Med. 1942 Jan 1;75(1):49–64. doi: 10.1084/jem.75.1.49. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Huang R. T., Rott R., Klenk H. D. Influenza viruses cause hemolysis and fusion of cells. Virology. 1981 Apr 15;110(1):243–247. doi: 10.1016/0042-6822(81)90030-1. [DOI] [PubMed] [Google Scholar]
  8. Jou W. M., Verhoeyen M., Devos R., Saman E., Fang R., Huylebroeck D., Fiers W., Threlfall G., Barber C., Carey N. Complete structure of the hemagglutinin gene from the human influenza A/Victoria/3/75 (H3N2) strain as determined from cloned DNA. Cell. 1980 Mar;19(3):683–696. doi: 10.1016/s0092-8674(80)80045-6. [DOI] [PubMed] [Google Scholar]
  9. Koshland D., Botstein D. Secretion of beta-lactamase requires the carboxy end of the protein. Cell. 1980 Jul;20(3):749–760. doi: 10.1016/0092-8674(80)90321-9. [DOI] [PubMed] [Google Scholar]
  10. Koszinowski U. H., Allen H., Gething M. J., Waterfield M. D., Klenk H. D. Recognition of viral glycoproteins by influenza A-specific cross-reactive cytolytic T lymphocytes. J Exp Med. 1980 Apr 1;151(4):945–958. doi: 10.1084/jem.151.4.945. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Krueger J. G., Wang E., Goldberg A. R. Evidence that the src gene product of Rous sarcoma virus is membrane associated. Virology. 1980 Feb;101(1):25–40. doi: 10.1016/0042-6822(80)90480-8. [DOI] [PubMed] [Google Scholar]
  12. Lai C. J., Markoff L. J., Zimmerman S., Cohen B., Berndt J. A., Chanock R. M. Cloning DNA sequences from influenza viral RNA segments. Proc Natl Acad Sci U S A. 1980 Jan;77(1):210–214. doi: 10.1073/pnas.77.1.210. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Laver W. G., Valentine R. C. Morphology of the isolated hemagglutinin and neuraminidase subunits of influenza virus. Virology. 1969 May;38(1):105–119. doi: 10.1016/0042-6822(69)90132-9. [DOI] [PubMed] [Google Scholar]
  14. Lazarowitz S. G., Choppin P. W. Enhancement of the infectivity of influenza A and B viruses by proteolytic cleavage of the hemagglutinin polypeptide. Virology. 1975 Dec;68(2):440–454. doi: 10.1016/0042-6822(75)90285-8. [DOI] [PubMed] [Google Scholar]
  15. Lohmeyer J., Klenk H. D. A mutant of influenza virus with a temperature-sensitive defect in the posttranslational processing of the hemagglutinin. Virology. 1979 Feb;93(1):134–145. doi: 10.1016/0042-6822(79)90282-4. [DOI] [PubMed] [Google Scholar]
  16. Matlin K. S., Reggio H., Helenius A., Simons K. Infectious entry pathway of influenza virus in a canine kidney cell line. J Cell Biol. 1981 Dec;91(3 Pt 1):601–613. doi: 10.1083/jcb.91.3.601. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Maxam A. M., Gilbert W. Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980;65(1):499–560. doi: 10.1016/s0076-6879(80)65059-9. [DOI] [PubMed] [Google Scholar]
  18. Mosmann T. R., Williamson A. R. Structural mutations in a mouse immunoglobulin light chain resulting in failure to be secreted. Cell. 1980 Jun;20(2):283–292. doi: 10.1016/0092-8674(80)90614-5. [DOI] [PubMed] [Google Scholar]
  19. Nakamura K., Compans R. W. Glycopeptide components of influenza viral glycoproteins. Virology. 1978 May 15;86(2):432–442. doi: 10.1016/0042-6822(78)90083-1. [DOI] [PubMed] [Google Scholar]
  20. Ploegh H. L., Orr H. T., Strominger J. L. Molecular cloning of a human histocompatibility antigen cDNA fragment. Proc Natl Acad Sci U S A. 1980 Oct;77(10):6081–6085. doi: 10.1073/pnas.77.10.6081. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Porter A. G., Barber C., Carey N. H., Hallewell R. A., Threlfall G., Emtage J. S. Complete nucleotide sequence of an influenza virus haemagglutinin gene from cloned DNA. Nature. 1979 Nov 29;282(5738):471–477. doi: 10.1038/282471a0. [DOI] [PubMed] [Google Scholar]
  22. Rogers J., Early P., Carter C., Calame K., Bond M., Hood L., Wall R. Two mRNAs with different 3' ends encode membrane-bound and secreted forms of immunoglobulin mu chain. Cell. 1980 Jun;20(2):303–312. doi: 10.1016/0092-8674(80)90616-9. [DOI] [PubMed] [Google Scholar]
  23. Sabatini D. D., Kreibich G., Morimoto T., Adesnik M. Mechanisms for the incorporation of proteins in membranes and organelles. J Cell Biol. 1982 Jan;92(1):1–22. doi: 10.1083/jcb.92.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Sasavage N. L., Smith M., Gillam S., Woychik R. P., Rottman F. M. Variation in the polyadenylylation site of bovine prolactin mRNA. Proc Natl Acad Sci U S A. 1982 Jan;79(2):223–227. doi: 10.1073/pnas.79.2.223. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Schwarz R. T., Schmidt M. F., Anwer U., Klenk H. D. Carbohydrates of influenza virus. I. Glycopeptides derived from viral glycoproteins after labeling with radioactive sugars. J Virol. 1977 Aug;23(2):217–226. doi: 10.1128/jvi.23.2.217-226.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Sveda M. M., Lai C. J. Functional expression in primate cells of cloned DNA coding for the hemagglutinin surface glycoprotein of influenza virus. Proc Natl Acad Sci U S A. 1981 Sep;78(9):5488–5492. doi: 10.1073/pnas.78.9.5488. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Sveda M. M., Markoff L. J., Lai C. J. Cell surface expression of the influenza virus hemagglutinin requires the hydrophobic carboxy-terminal sequences. Cell. 1982 Sep;30(2):649–656. doi: 10.1016/0092-8674(82)90261-6. [DOI] [PubMed] [Google Scholar]
  28. Wehland J., Willingham M. C., Gallo M. G., Pastan I. The morphologic pathway of exocytosis of the vesicular stomatitis virus G protein in cultured fibroblasts. Cell. 1982 Apr;28(4):831–841. doi: 10.1016/0092-8674(82)90062-9. [DOI] [PubMed] [Google Scholar]
  29. Wiley D. C., Skehel J. J., Waterfield M. Evidence from studies with a cross-linking reagent that the haemagglutinin of influenza virus is a trimer. Virology. 1977 Jun 15;79(2):446–448. doi: 10.1016/0042-6822(77)90371-3. [DOI] [PubMed] [Google Scholar]
  30. Zilberstein A., Snider M. D., Porter M., Lodish H. F. Mutants of vesicular stomatitis virus blocked at different stages in maturation of the viral glycoprotein. Cell. 1980 Sep;21(2):417–427. doi: 10.1016/0092-8674(80)90478-x. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Virology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES