Structure of Adenovirus Type 21 Knob in Complex with CD46 Reveals Key Differences in Receptor Contacts among Species B Adenoviruses

ABSTRACT The complement regulation protein CD46 is the primary attachment receptor for most species B adenoviruses (Ads). However, significant variability exists in sequence and structure among species B Ads in the CD46-binding regions, correlating with differences in affinity. Here, we report a structure-function analysis of the interaction of the species B Ad21 knob with the two N-terminal repeats SCR1 and SCR2 of CD46, CD46-D2. We have determined the structures of the Ad21 knob in its unliganded form as well as in complex with CD46-D2, and we compare the interactions with those observed for the Ad11 knob-CD46-D2 complex. Surface plasmon resonance measurements demonstrate that the affinity of Ad21 knobs for CD46-D2 is 22-fold lower than that of the Ad11 knob. The superposition of the Ad21 and Ad11 knob structures in complex with CD46-D2 reveals a substantially different binding mode, providing an explanation for the weaker binding affinity of the Ad21 knob for its receptor. A critical difference in both complex structures is that a key interaction point, the DG loop, protrudes more in the Ad21 knob than in the Ad11 knob. Therefore, the protruding DG loop does not allow CD46-D2 to approach the core of the Ad21 knob as closely as in the Ad11 knob-CD46-D2 complex. In addition, the engagement of CD46-D2 induces a conformational change in the DG loop in the Ad21 knob but not in the Ad11 knob. Our results contribute to a more profound understanding of the CD46-binding mechanism of species B Ads and have relevance for the design of more efficient gene delivery vectors.

[1]  T. Stehle,et al.  Specificity switching in virus–receptor complexes , 2009, Current Opinion in Structural Biology.

[2]  Marko Marttila,et al.  An Arginine Switch in the Species B Adenovirus Knob Determines High-Affinity Engagement of Cellular Receptor CD46 , 2008, Journal of Virology.

[3]  G. Nemerow,et al.  Conservation of fiber structure and CD46 usage by subgroup B2 adenoviruses. , 2008, Virology.

[4]  G. Nemerow,et al.  Structural Variations in Species B Adenovirus Fibers Impact CD46 Association , 2008, Journal of Virology.

[5]  D. Metzgar,et al.  Abrupt emergence of diverse species B adenoviruses at US military recruit training centers. , 2007, The Journal of infectious diseases.

[6]  C. Verlinde,et al.  Identification of CD46 Binding Sites within the Adenovirus Serotype 35 Fiber Knob , 2007, Journal of Virology.

[7]  E. Delwart,et al.  New Adenovirus Species Found in a Patient Presenting with Gastroenteritis , 2007, Journal of Virology.

[8]  T. Stehle,et al.  Adenovirus type 11 binding alters the conformation of its receptor CD46 , 2007, Nature Structural &Molecular Biology.

[9]  Olivier Billet,et al.  Structural and Mutational Analysis of Human Ad37 and Canine Adenovirus 2 Fiber Heads in Complex with the D1 Domain of Coxsackie and Adenovirus Receptor* , 2006, Journal of Biological Chemistry.

[10]  A. Segerman,et al.  The Arg279Gln Substitution in the Adenovirus Type 11p (Ad11p) Fiber Knob Abolishes EDTA-Resistant Binding to A549 and CHO-CD46 Cells, Converting the Phenotype to That of Ad7p , 2006, Journal of Virology.

[11]  J. Atkinson,et al.  CD46 Is a Cellular Receptor for All Species B Adenoviruses except Types 3 and 7 , 2005, Journal of Virology.

[12]  C. Rooney,et al.  Adenovirus as an emerging pathogen in immunocompromised patients , 2005, British journal of haematology.

[13]  S. S. Oliveira,et al.  Characterization of species B adenoviruses isolated from fecal specimens taken from poliomyelitis-suspected cases. , 2004, Journal of clinical virology : the official publication of the Pan American Society for Clinical Virology.

[14]  R. Cattaneo.  Four Viruses, Two Bacteria, and One Receptor: Membrane Cofactor Protein (CD46) as Pathogens' Magnet , 2004, Journal of Virology.

[15]  A. Gaggar,et al.  CD46 is a cellular receptor for group B adenoviruses , 2003, Nature Medicine.

[16]  J. Atkinson,et al.  Adenovirus Type 11 Uses CD46 as a Cellular Receptor , 2003, Journal of Virology.

[17]  Rodrigo Lopez,et al.  Multiple sequence alignment with the Clustal series of programs , 2003, Nucleic Acids Res..

[18]  U. Greber,et al.  Adenovirus triggers macropinocytosis and endosomal leakage together with its clathrin-mediated uptake , 2002, The Journal of cell biology.

[19]  P. Quax,et al.  Exploiting the Natural Diversity in Adenovirus Tropism for Therapy and Prevention of Disease , 2002, Journal of Virology.

[20]  Randy J. Read,et al.  Pushing the boundaries of molecular replacement with maximum likelihood. , 2001, Acta crystallographica. Section D, Biological crystallography.

[21]  S. Cusack,et al.  Structure of the fiber head of Ad3, a non-CAR-binding serotype of adenovirus. , 2001, Virology.

[22]  Qin Chen,et al.  Adenovirus is a key pathogen in hemorrhagic cystitis associated with bone marrow transplantation. , 2001, Clinical infectious diseases : an official publication of the Infectious Diseases Society of America.

[23]  G. Stamatoyannopoulos,et al.  Efficient Gene Transfer into Human CD34+ Cells by a Retargeted Adenovirus Vector , 2000, Journal of Virology.

[24]  A. Kidd,et al.  Adenovirus Type 37 Uses Sialic Acid as a Cellular Receptor , 2000, Journal of Virology.

[25]  M. Malnati,et al.  CD46 Is a Cellular Receptor for Human Herpesvirus 6 , 1999, Cell.

[26]  M. Bewley,et al.  Structural analysis of the mechanism of adenovirus binding to its human cellular receptor, CAR. , 1999, Science.

[27]  R. Ravelli,et al.  Adhesion mechanism of human β2‐glycoprotein I to phospholipids based on its crystal structure , 1999, The EMBO journal.

[28]  T. Stehle,et al.  Crystal structure of two CD46 domains reveals an extended measles virus‐binding surface , 1999, The EMBO journal.

[29]  D. Brough,et al.  The Coxsackievirus-Adenovirus Receptor Protein Can Function as a Cellular Attachment Protein for Adenovirus Serotypes from Subgroups A, C, D, E, and F , 1998, Journal of Virology.

[30]  R J Read,et al.  Crystallography & NMR system: A new software suite for macromolecular structure determination. , 1998, Acta crystallographica. Section D, Biological crystallography.

[31]  J. Atkinson,et al.  Membrane cofactor protein (MCP or CD46) is a cellular pilus receptor for pathogenic Neisseria , 1997, Molecular microbiology.

[32]  G. Murshudov,et al.  Refinement of macromolecular structures by the maximum-likelihood method. , 1997, Acta crystallographica. Section D, Biological crystallography.

[33]  Wolfgang Kabsch,et al.  Automatic processing of rotation diffraction data from crystals of initially unknown symmetry and cell constants , 1993 .

[34]  Ari Helenius,et al.  Stepwise dismantling of adenovirus 2 during entry into cells , 1993, Cell.

[35]  Christopher D. Richardson,et al.  The human CD46 molecule is a receptor for measles virus (Edmonston strain) , 1993, Cell.

[36]  H. Cushing,et al.  Severe adenoviral nephritis following bone marrow transplantation: successful treatment with intravenous ribavirin. , 1993, Bone marrow transplantation.

[37]  D. Gerlier,et al.  Human membrane cofactor protein (CD46) acts as a cellular receptor for measles virus , 1993, Journal of virology.

[38]  G. Nemerow,et al.  Integrins α v β 3 and α v β 5 promote adenovirus internalization but not virus attachment , 1993, Cell.

[39]  J. D. de Jong,et al.  Molecular epidemiology of adenovirus type 21 in the Netherlands and the Federal Republic of Germany from 1960 to 1985 , 1986, Journal of clinical microbiology.

[40]  J. Atkinson,et al.  Purification and characterization of a membrane protein (gp45-70) that is a cofactor for cleavage of C3b and C4b , 1986, The Journal of experimental medicine.

[41]  S. Darougar,et al.  Adenovirus type 21 keratoconjunctivitis. , 1978, The British journal of ophthalmology.

[42]  W. Delano.  The PyMOL Molecular Graphics System , 2002 .

[43]  J. Atkinson,et al.  Membrane cofactor protein (MCP or CD46): newest member of the regulators of complement activation gene cluster. , 1991, Annual review of immunology.

[44]  Randy J Read,et al.  Electronic Reprint Biological Crystallography Phenix: Building New Software for Automated Crystallographic Structure Determination Biological Crystallography Phenix: Building New Software for Automated Crystallographic Structure Determination , 2022 .