Mutational mapping of UL130 of human cytomegalovirus defines peptide motifs within the C-terminal third as essential for endothelial cell infection.
The UL130 gene is one of the major determinants of endothelial cell (EC) tropism of human cytomegalovirus (HCMV). In order to define functionally important peptides within this protein, we have performed a charge-cluster-to-alanine (CCTA) mutational scanning of UL130 in the genetic background of a bacterial artificial chromosome-cloned endotheliotropic HCMV strain. A total of 10 charge clusters were defined, and in each of them two or three charged amino acids were replaced with alanines. While the six N-terminal clusters were phenotypically irrelevant, mutation of the four C-terminal clusters each caused a reduction of EC tropism. The importance of this protein domain was further emphasized by the fact that the C-terminal pentapeptide PNLIV was essential for infection of ECs, and the cell tropism could not be rescued by a scrambled version of this sequence. We conclude that the C terminus of the UL130 protein serves an important function for infection of ECs by HCMV. This makes UL130 a promising molecular target for antiviral strategies, e.g., the development of antiviral peptides.
Schuessler
Andrea
A
Institut für Medizinische Virologie, Eberhard-Karls-Universität, Tübingen, Germany.
Sampaio
Kerstin Laib
KL
Scrivano
Laura
L
Sinzger
Christian
C
D020816:Amino Acid Motifs; D002478:Cells, Cultured; D003587:Cytomegalovirus / Q000235:genetics / Q000472:pathogenicity / Q000502:physiology*; D004252:DNA Mutational Analysis; D004279:DNA, Viral / Q000235:genetics*; D042783:Endothelial Cells / Q000821:virology*; D006801:Humans; D008562:Membrane Glycoproteins / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D017434:Protein Structure, Tertiary; D014759:Viral Envelope Proteins / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D056189:Viral Tropism; D037521:Virulence Factors / Q000235:genetics / Q000502:physiology*
D004279:DNA, Viral; D008562:Membrane Glycoproteins; C502890:UL130 protein, human cytomegalovirus; D014759:Viral Envelope Proteins; D037521:Virulence Factors