SUMO-1 modification of the major immediate-early (IE) 1 and 2 proteins of human cytomegalovirus is regulated by different mechanisms and modulates the intracellular localization of the IE1, but not IE2, protein.
We have previously shown that two proteins with apparent molecular masses of 91- and 102-kDa (p91 and p102, respectively) in human cytomegalovirus (HCMV)-infected cells are antigenically and structurally related to the major immediate-early (IE) 1 and 2 proteins (IE1p68 and IE2p80, respectively) of HCMV, respectively. In this study, we extended the characterization of p91 and p102 and our results were as follows; (i) Lysine at amino acid position 450 in IE1p68 and at amino acid position 175 or 180 in IE2p80, to which SUMO-1 has been shown to be covalently linked, are required for production of p91 and p102, respectively. (ii) A reversal of cycloheximide (CH) block in the presence of actinomycin D imposed at the time of infection inhibited production of p91, but not p102. (iii) The steady-state levels of p91, but not p102, were remarkably decreased by treatment with proteasome inhibitor MG132, but coincubation with CH inhibited this decrease of p91. (iv) Cell fractionation by differential detergent extraction demonstrated that p91 is preferentially found in detergent-insoluble fraction, although p102 as well as IE1p68 and IE2p80 distributes into all fractions. These results demonstrate that p91 and p102 correspond to SUMO-1-modified IE1p68 and IE2p80, respectively, that the production and degradation of SUMO-1-modified IE1p68 is regulated by mechanisms different from those of SUMO-1-modified IE2p80, and that SUMO-1 modification modulates the intracellular localization of IE1p68, but not IE2p80.
Sadanari
H
H
Department of Microbiology, Faculty of Pharmaceutical Sciences, Hokuriku University, Kanazawa, Japan. h-sadanari@hokuriku-u.ac.jp
Yamada
R
R
Ohnishi
K
K
Matsubara
K
K
Tanaka
J
J
D002460:Cell Line; D003587:Cytomegalovirus / Q000378:metabolism*; D006801:Humans; D017874:Immediate-Early Proteins / Q000737:chemistry / Q000378:metabolism*; D008970:Molecular Weight; D025842:SUMO-1 Protein / Q000502:physiology*; D015534:Trans-Activators / Q000737:chemistry / Q000378:metabolism*; D014764:Viral Proteins / Q000737:chemistry / Q000378:metabolism*
C088645:IE1 protein, cytomegalovirus; C069186:IE2 protein, Cytomegalovirus; D017874:Immediate-Early Proteins; D025842:SUMO-1 Protein; D015534:Trans-Activators; D014764:Viral Proteins