Alphaherpesvirinae and Gammaherpesvirinae glycoprotein L and CMV UL130 originate from chemokines.

2.5 MB·English

by  Malkowska, Maja

Malkowskaetal.VirologyJournal2013,10:1
http://www.virologyj.com/content/10/1/1
HYPOTHESIS Open Access
Alphaherpesvirinae and Gammaherpesvirinae
glycoprotein L and CMV UL130 originate from
chemokines
Maja Malkowska1, Katarzyna Kokoszynska1, Magdalena Dymecka1,2, Leszek Rychlewski2 and Lucjan S Wyrwicz1*
Abstract
Herpesviridae is a large family of DNA viruses divided intothree subfamilies: Alpha-,Beta- and Gammaherpesvirinae.
The process of herpesvirus transmissionis mediated bya range of proteins, one ofwhich is glycoproteinL (gL).
Based onour analysis ofthe solved structures of HSV2 and EBV gH/gL complexes,we propose that
Alphaherpesvirinae and Gammaherpesvirinae glycoproteinL and Betaherpesvirinae UL130 originate from chemokines.
Herpes simplex virus type 2gL and humancytomegalovirus homolog (UL130) adopt a novelC chemokine-like fold,
whileEpstein-Barr virus gL mimics a CCchemokine structure. Hence, it is possible that gL interface with specific
chemokine receptors during the transmission of Herpesviridae. We conclude that the further understanding of the
function of viral chemokine-like proteins inHerpesviridae infection maylead to development ofnovelprophylactic
and therapeutic treatment.
Keywords: Herpesviridae,Glycoprotein L,GL, UL130, Chemokines, HCMV, HSV2,EBV
Background studies on gH/gL biology have been carried out, there is
Asa family of dsDNAviruses, Herpesviridae are known to still no unequivocal evidence whether this complex plays
infectvertebrates,includinghumans,andcauseprogression role in the recognition of specific cells (tissues) or directly
of various contagious diseases, such as orofacial herpes takespartinmembranefusion[5].
(HSV1), genital herpes (HSV2), chickenpox and shingles Thechemokinefamilyhasbeencreatedbasedonstruc-
(VZV), opportunistic infections (CMV), Kaposi’s sarcoma tural similarity. The characteristics features are the Greek
(KSHV) and mononucleosis (EBV) [1]. Members of the key shape and conserved cysteine residues forming
Herpesviridaefamilyhavebeenclassifiedintothreedistinct SS-bonds and stabilizing the structure. Chemokine family
subfamilies: Alpha- (HSV1, HSV2, VZV); Beta- (CMV, differs in terms of sequence similarity, which vary from
HHV-6,HHV-7);Gammaherpesvirinae(EBV,KSHV). less than 20% to over 90% [6]. Based on the position of
The cell-to-cell transmission of virions is one of limiting conservedcysteineschemokine classificationdividesthem
steps in spread of infection. The targeting of a specific tis- intofoursub-families(CC,CXC,CandCX C).
3
sue for infection is based on the recognition of definite Chemokine mimicry is a common phenomenon among
entry receptors [2]. The virus used as a model of herpetic herpesvirus, poxvirus and retrovirus families [7-9]. So far,
infectionisHSV1.Initsgenome,11outof80genesencode over 30 chemokine and chemokine receptor mimics have
outer membrane glycoproteins. According to previous been described. Except for HCMV UL130 and HSV2 gL
studies, glycoprotein D (gD), glycoprotein B (gB) and the no other viral proteins have been designated as C type
complex of glycoproteins H and L (gH/gL) are curial pro- chemokinehomologs[10-12].
teins for cell-to-cell fusion [3,4]. Although numerous
Presentationofthehypothesis
*Correspondence:[email protected] In our previous report, based on bioinformatic analysis of
1LaboratoryofBioinformaticsandBiostatistics,MariaSklodowska-Curie protein families, we concluded the possibility of a distant
MemorialCancerCenterandInstituteofOncology,WKRoentgena5,02-781, homology between Herpesviridae glycoprotein L (gL)
Warsaw,Poland
Fulllistofauthorinformationisavailableattheendofthearticle and chemokines [12]. We also remarked that human
©2013Malkowskaetal.;licenseeBioMedCentralLtd.ThisisanOpenAccessarticledistributedunderthetermsofthe
CreativeCommonsAttributionLicense(http://creativecommons.org/licenses/by/2.0),whichpermitsunrestricteduse,
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Cytomegalovirus (HCMV) contains an additional gene
(UL130) which might encode an orthologous chemokine-
like protein. This study was followed by crystallographic
determination of the structure of two gH/gL complexes.
Chowdary andco-workerscrystallographically determined
thestructureoftheHSV2(Herpessimplexvirus2)gH/gL
complex,whilstMatsuuraetal.describedthecrystalstruc-
tureofanEpstein-Barrvirus(EBV)complex[13,14].
Although not noticed in the original reports by
Chowdary et al. and Matsuura et al., it is important to
note that the topologies of both solved glycoproteins are
highly concordantwith thehomologymodels [12].
We hypothesize that the betaherpevirus UL130 as well
as alpha- and gammaherpesvirus gL proteins exhibit
structural similarity with chemokines and may interact
with theirreceptors.
The structural comparison between EBV gL and
CC-type chemokines leaves no doubt that it adopts a
CC chemokine-like fold. Figure 1 shows the superim- Figure1SuperimposednativestructuresofC-Cmotif
chemokine5(green;PDBentry:1U4LA)andEBVglycoprotein
posed structures of a CC-type chemokine (human C-C
L(cyan;PDBentry3PHFB).Thecysteineresiduesaremarkedin
motif chemokine 5; Protein Data Bank entry: 1U4L_A)
red(C-Cmotifchemokine5)andmagenta(gL).
and the native structure of EBV glycoprotein L (Protein
Data Bank entry 3PHF_B) which exhibit a root mean
square deviation (RMSD) for C-alpha atoms of 1.23 Å UL130 to C type chemokine subfamily (Figure 5). Despite
(analysis performed for atoms forming the curved beta- of the loss of 2nd and 4th cytosine the chemotactic activity
sheet). The conserved topology is a result of the pres- might be preserved and requires further investigation as it
ence of two cysteine bridges (see Figure 1). Furthermore, might explain the mechanism of UL130 functioning in
the putative interaction surface of EBV’s glycoprotein L EndothelialCellinfection[17].
with chemokines is also preserved (for the conserved Mutagenesis of interleukine-8 showed that perturbation
pattern of hydrophobic amino acids see Figure 2). The of the cysteine bridge between 2nd and 4th motif element
major difference is a result of the altered angle between hadsmalleffectsonitsfunctionwhereas themodification
the helix and the curved beta-sheet, which is to be of the disulfide between 1st and 3rd cysteine dramatically
expected, since the helix in chemokines is involved in reduced potency [18]. This fact supports our hypothesis
dimerizationduringreceptor recognition, whilstgL lacks
the ability to dimerize. A further difference can be
seen in the presence of two additional helices at the
C-terminus of glycoprotein L – 15 and 6 residues long
respectively,separated byalinkofonlytwo aminoacids.
Comparison of cysteine positions in CXC motif in
interleukine-8 (IL-8) and lymphotactin (C type chemo-
kine) has shown the conservation of 1nd and 4th cysteine
(Figure3),whereasincaseofIL-8andgLfrom HSV2the
1st and 3rd cysteines are conserved (Figure 4). Despite the
non-conserved cysteine position, based on structural
similarity, we propose that gL HSV2 shall be classified as
Ctypechemokine-likeprotein.
Due to synteny, rat cytomegalovirus r131 gene encoding
a proinflammatory CC chemokine-like protein has been
recognized as HCMV UL130 homolog [15,16]. Sequence
identity and similarity between r131 and UL130 is 19.1% Figure2SuperimposednativestructuresoftheCC-type
and 41.1% respectively (calculations were performed with chemokineeotaxin-2(green;PDBentry:1EIGA)andEBV
PAM250scoringmatrixforglobalalignment).Theposition glycoproteinL(light-green;PDBentry3PHFB).Thehydrophobic
ofconservedcysteines(1stand3rdcysteineofcharacteristic surfacesaremarkedinblue(eotaxin-2)andlight-blue(gL),cysteine
residuesinmagenta(eotaxin-2)andlight-pink(gL).
CXC type chemokine motif) leads to the assignment of
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Figure3Multiplesequencealignmentoflymphotactin(XCL1)andinterleukin-8(IL-8).Thecorrespondingsequenceswerelabeledwith
theirGenBankentries(denotedbytheGenBankidentifier-gi)andorganismoforigin.Thenumbersinbracketsrefertothepositionsofthe
presentedsequencefragments.Theobserved(ProteinDataBankentries1J8I_AforXCL1and1IL8_AforIL-8)andpredicted(Psipred)secondary
structureelementsarecodedwithletters(H-a-helix,G-3 helix,E-b-strands).DisulphidebridgesinIL-8hasbeenmarkedwitharrows
10
andnumbered.
that partial conservation of cysteines in UL130 does not [22].AlignmentfiguresweremadewiththeuseofBioEdit
necessarilyimplyalackofchemotacticactivity. version 7.1.3 alignment editor [23]. Residue conservation
The speculation concerning the co-evolution of the two werepresentedinalignmentwiththefollowingdefaultfor
interactingglycoproteins[14]causesustopresumethatthe thesoftwarescheme:positivelychargedhighlightedinred;
most likely cause of such concordance is that glycoprotein negatively charged in blue; hydrophobic in dark green;
L arose asa result oflateral gene transfer from thehost.It tryptophan in light green; polar uncharged and proline in
is very likely that early in the evolution of Herpesviridae, gray; histidine in magenta; glycine in yellow; alanine,
glycoprotein L acted predominantly as a virus encoded phenylalanine and tyrosine in cyan; cysteine in maroon.
chemokine (compare viral chemokines: UL146, UL147, Modeller9wasusedtocreatehomologymodels[24].The
UL152 encoded by HCMV [19]), which later co-evolved structures were visualized with PyMOL v. 0.99rc6 which
withtheothermembraneviralgenes. alsoenablesRMSDcalculations[25].
In presented analysis we have used a typical homology
modeling workbench. The sets of homologs from the NR Testingthehypothesis
database (NCBI; National Center for Biotechnology) were Verylowsequencesimilaritybetweenvariouschemokines
createdusingPSI-BLAST[20]search methodandaligned and gL prevents any phylogenetic studies of the origin of
with ClustalW [21]. Secondary structure predictions for these protein families from being carried out, and we are
selected proteins were calculated using PSIPRED method unable to justify whether the different topology types
Figure4MultiplesequencealignmentofglycoproteinL(foundinAlphaherpesvirinae)andinterleukin-8(IL-8).Thecorresponding
sequenceswerelabeledwiththeirGenBankentries(denotedbytheGenBankidentifier-gi)andorganismoforigin.Thenumbersinbrackets
refertothepositionsofthepresentedsequencefragments.Theobserved(ProteinDataBankentries3M1C_BforgLand1IL8_AforIL-8)and
predicted(Psipred)secondarystructureelementsarecodedwithletters(H-a-helix,G-3 helix,E-b-strands).DisulphidebridgesinIL-8has
10
beenmarkedwitharrowsandnumbered.
Malkowskaetal.VirologyJournal2013,10:1 Page4of5
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Figure5MultiplesequencealignmentofUL130(foundinBetaherpesvirinae)andinterleukin-8(IL-8).Thecorrespondingsequenceswere
labeledwiththeirGenBankentries(denotedbytheGenBankidentifier-gi)andorganismoforigin.Thenumbersinbracketsrefertothe
positionsofthepresentedsequencefragments.Theobserved(ProteinDataBankentry1IL8_AforIL-8)andpredicted(Psipred)secondary
structureelementsarecodedwithletters(H-a-helix,G-3 helix,E-b-strands).DisulphidebridgesinIL-8hasbeenmarkedwitharrows
10
andnumbered.
(CC for Gammaherpesvirinae, C for the Alpha group and Competinginterests
HCMV) come from a single event in the early divergence Theauthorsdeclarethattheyhavenocompetinginterests.
of Herpesviridae, or rather arose from two independent
Authors’contributions
eventsoflateralgenetransfer.
MM,KK,LSWparticipatedinthedesignofthestudy.MM,KK,MD,LRand
Discussed here glycoproteins are present in viral enve- LSWcarriedoutthebioinformaticsanalysis.MM,KK,LSWwrotethe
lope and mediate virus entry to host cells. Their mech- manuscript.Allauthorsreadandapprovedthefinalmanuscript.
anism and function as chemokine-like proteins however,
Acknowledgements
remains unknown. There is a potential for further ana-
ThisworkwassupportedbythePolishNationalCentreforResearchand
lysis by experimental characterization of specific chemo- Development(NCBiR)INNOTECHProgramme(IN1/159417).
tactic activities of gL and UL130 proteins. Necessary
Authordetails
step would be in vivo expression during viral persistence
1LaboratoryofBioinformaticsandBiostatistics,MariaSklodowska-Curie
andverification oftheirrelease from cells. Toestablishif MemorialCancerCenterandInstituteofOncology,WKRoentgena5,02-781,
theseproteinsarefunctionalchemokinesandwhichcells Warsaw,Poland.2BioInfoBankInstitute,Limanowskiego24A,60-744,Poznań,
Poland.
are their targets in vivo migration assays shall be per-
formed. Site-directed mutagenesis would help to deter- Received:6April2012Accepted:18December2012
mine functional domains that are critical for migration. Published:2January2013
Ligand-binding assays could indicate binding affinity to
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doi:10.1186/1743-422X-10-1
Citethisarticleas:Malkowskaetal.:Alphaherpesvirinaeand
GammaherpesvirinaeglycoproteinLandCMVUL130originatefrom
chemokines.VirologyJournal201310:1.
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