Human cytomegalovirus major immediate early 1 protein targets host chromosomes by docking to the acidic pocket on the nucleosome surface.
The 72-kDa immediate early 1 (IE1) protein encoded by human cytomegalovirus (hCMV) is a nuclearly localized promiscuous regulator of viral and cellular transcription. IE1 has long been known to associate with host mitotic chromatin, yet the mechanisms underlying this interaction have not been specified. In this study, we identify the cellular chromosome receptor for IE1. We demonstrate that the viral protein targets human nucleosomes by directly binding to core histones in a nucleic acid-independent manner. IE1 exhibits two separable histone-interacting regions with differential binding specificities for H2A-H2B and H3-H4. The H2A-H2B binding region was mapped to an evolutionarily conserved 10-amino-acid motif within the chromatin-tethering domain (CTD) of IE1. Results from experimental approaches combined with molecular modeling indicate that the IE1 CTD adopts a β-hairpin structure, docking with the acidic pocket formed by H2A-H2B on the nucleosome surface. IE1 binds to the acidic pocket in a way similar to that of the latency-associated nuclear antigen (LANA) of the Kaposi's sarcoma-associated herpesvirus. Consequently, the IE1 and LANA CTDs compete for binding to nucleosome cores and chromatin. Our work elucidates in detail how a key viral regulator is anchored to human chromosomes and identifies the nucleosomal acidic pocket as a joint target of proteins from distantly related viruses. Based on the striking similarities between the IE1 and LANA CTDs and the fact that nucleosome targeting by IE1 is dispensable for productive replication even in "clinical" strains of hCMV, we speculate that the two viral proteins may serve analogous functions during latency of their respective viruses.
Institute for Medical Microbiology and Hygiene, University of Regensburg, Regensburg, Germany.
Katrin
K
Mücke
Christina
C
Paulus
Katharina
K
Bernhardt
Katrin
K
Gerrer
Kathrin
K
Schön
Alina
A
Fink
Eva-Maria
EM
Sauer
Alexandra
A
Asbach-Nitzsche
Thomas
T
Harwardt
Bärbel
B
Kieninger
Werner
W
Kremer
Hans Robert
HR
Kalbitzer
Michael
M
Nevels
D000595:Amino Acid Sequence; D001665:Binding Sites; D002877:Chromosomes, Human / Q000235:genetics / Q000378:metabolism / Q000821:virology*; D003587:Cytomegalovirus / Q000235:genetics / Q000378:metabolism*; D003586:Cytomegalovirus Infections / Q000235:genetics / Q000378:metabolism / Q000821:virology*; D006657:Histones / Q000235:genetics / Q000378:metabolism; D006801:Humans; D017874:Immediate-Early Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009707:Nucleosomes / Q000737:chemistry / Q000378:metabolism* / Q000821:virology; D011485:Protein Binding; D017434:Protein Structure, Tertiary
D006657:Histones; C088645:IE1 protein, cytomegalovirus; D017874:Immediate-Early Proteins; D009707:Nucleosomes