Ultrastructural and immunocytochemical evidences of core-particle formation in the methylotrophic Pichia pastoris yeast when expressing HCV structural proteins (core-E1)

Particulate antigens of the Hepatitis C virus (HCV) are reported for the first time by transmission electron microscopy in Pichia pastoris. The yeast was cloned to express the first 339 NH2-terminal amino acids of the HCV polyprotein (C-E1.339 polypeptide). The C-E1.339 polypeptide covers the putati...

Full description

Saved in:
Bibliographic Details
Published inTissue & cell Vol. 31; no. 2; pp. 117 - 125
Main Authors Falcón, V, García, C, de la Rosa, M C, Menéndez, I, Seoane, J, Grillo, J M
Format Journal Article
LanguageEnglish
Published Scotland 01.04.1999
Subjects
Online AccessGet full text

Cover

Loading…
Abstract Particulate antigens of the Hepatitis C virus (HCV) are reported for the first time by transmission electron microscopy in Pichia pastoris. The yeast was cloned to express the first 339 NH2-terminal amino acids of the HCV polyprotein (C-E1.339 polypeptide). The C-E1.339 polypeptide covers the putative 191 aa of the core protein (aa 1-191) and 148 aa of the E1 envelope antigen (aa 192-339). Virus-like particles (VLP) with diameters ranging from 20 nm to 30 nm were specifically observed in those cells expressing the HCV polyprotein. The VLP appeared along the membrane of the endoplasmic reticulum, but were fundamentally localized in vacuoles, either free or inside autophagic bodies. Clustered particles, chains of particles, high-density reticular structures, and crystalloid bodies were also detected, the last one being an orderly arrangement of particles with 20 nm diameters. The crystal-associated particles are well differentiated from the intracellular VLP because of their uniform size and shape. We argue that membrane components are retained in the architecture of the VLP, conferring to this particle certain heterogeneity. Both kinds of particles, the VLP formed after treatment with NP-40 and the crystal-associated particles, were core protein-positives. Whether they reflect mature HCV nucleocapsid or intermediary states in the viral nucleocapsid morphogenesis remains unknown. We conclude that, like mammalian cell lines, the P. pastoris yeast could be an appropriate host for the analysis of HCV polyprotein processing and, eventually, virus assembly.
AbstractList Particulate antigens of the Hepatitis C virus (HCV) are reported for the first time by transmission electron microscopy in Pichia pastoris. The yeast was cloned to express the first 339 NH2-terminal amino acids of the HCV polyprotein (C-E1.339 polypeptide). The C-E1.339 polypeptide covers the putative 191 aa of the core protein (aa 1-191) and 148 aa of the E1 envelope antigen (aa 192-339). Virus-like particles (VLP) with diameters ranging from 20 nm to 30 nm were specifically observed in those cells expressing the HCV polyprotein. The VLP appeared along the membrane of the endoplasmic reticulum, but were fundamentally localized in vacuoles, either free or inside autophagic bodies. Clustered particles, chains of particles, high-density reticular structures, and crystalloid bodies were also detected, the last one being an orderly arrangement of particles with 20 nm diameters. The crystal-associated particles are well differentiated from the intracellular VLP because of their uniform size and shape. We argue that membrane components are retained in the architecture of the VLP, conferring to this particle certain heterogeneity. Both kinds of particles, the VLP formed after treatment with NP-40 and the crystal-associated particles, were core protein-positives. Whether they reflect mature HCV nucleocapsid or intermediary states in the viral nucleocapsid morphogenesis remains unknown. We conclude that, like mammalian cell lines, the P. pastoris yeast could be an appropriate host for the analysis of HCV polyprotein processing and, eventually, virus assembly.
Particulate antigens of the Hepatitis C virus (HCV) are reported for the first time by transmission electron microscopy in Pichia pastoris. The yeast was cloned to express the first 339 NH sub(2)-terminal amino acids of the HCV polyprotein (C-E1.339 polypeptide). The C-E1.339 polypeptide covers the putative 191 aa of the core protein (aa 1-191) and 148 aa of the E1 envelope antigen (aa 192-339). Virus-like particles (VLP) with diameters ranging from 20 nm to 30 nm were specifically observed in those cells expressing the HCV polyprotein. The VLP appeared along the membrane of the endoplasmic reticulum, but were fundamentally localized in vacuoles, either free or inside autophagic bodies. Clustered particles, chains of particles, high-density reticular structures, and crystalloid bodies were also detected, the last one being an orderly arrangement of particles with 20 nm diameters. The crystal-associated particles are well differentiated from the intracellular VLP because of their uniform size and shape. We argue that membrane components are retained in the architecture of the VLP, conferring to this particle certain heterogeneity. Both kinds of particles, the VLP formed after treatment with NP-40 and the crystal-associated particles, were core protein-positives. Whether they reflect mature HCV nucleocapsid or intermediary states in the viral nucleocapsid morphogenesis remains unknown. We conclude that, like mammalian cell lines, the P. pastoris yeast could be an appropriate host for the analysis of HCV polyprotein processing and, eventually, virus assembly.
Author Menéndez, I
Falcón, V
García, C
Grillo, J M
Seoane, J
de la Rosa, M C
Author_xml – sequence: 1
  givenname: V
  surname: Falcón
  fullname: Falcón, V
  organization: Electron Microscopy Department, Center for Genetic Engineering and Biotechnology (CIGB), Havana, Cuba
– sequence: 2
  givenname: C
  surname: García
  fullname: García, C
– sequence: 3
  givenname: M C
  surname: de la Rosa
  fullname: de la Rosa, M C
– sequence: 4
  givenname: I
  surname: Menéndez
  fullname: Menéndez, I
– sequence: 5
  givenname: J
  surname: Seoane
  fullname: Seoane, J
– sequence: 6
  givenname: J M
  surname: Grillo
  fullname: Grillo, J M
BackLink https://www.ncbi.nlm.nih.gov/pubmed/10445295$$D View this record in MEDLINE/PubMed
BookMark eNqFkU1LxDAQhnNY8fvqUXISPXRNmqRpjrKoKwh6cL0uaTq1kTapSaruT_JfWvwAb55mGB7meZnZQzPnHSB0RMmcEsHPkzUwp0qpOSEsn6FdQjjJSloUO2gvxmdCiORUbqMdSjgXuRK76GPVpaBjCqNJY9Ad1q7Gtu9H580medNCb800hldbgzMQsW-w8QGyQYdJ2AFufOh1st5h63BqAfeQ2k3nU_BDaw2-t6a1Gg-TxQcb8QamDr-14DC8DwFitO4JLxeP-E-MIfgE1kV8-iW7pGcHaKvRXYTDn7qPVleXD4tldnt3fbO4uM2GnJUpY5xJoCA544LzpikrXhsqhAEjZCOUqqRkijEGOVEVK6igui4KXVFtNJ8Ot49OvvdOEV5GiGnd22ig67QDP8Z1oZSQgrJ_QSrziS3LCTz-Aceqh3o9BNvrsFn_foF9AuWmjdE
ContentType Journal Article
DBID CGR
CUY
CVF
ECM
EIF
NPM
7U9
H94
7X8
DOI 10.1054/tice.1999.0032
DatabaseName Medline
MEDLINE
MEDLINE (Ovid)
MEDLINE
MEDLINE
PubMed
Virology and AIDS Abstracts
AIDS and Cancer Research Abstracts
MEDLINE - Academic
DatabaseTitle MEDLINE
Medline Complete
MEDLINE with Full Text
PubMed
MEDLINE (Ovid)
AIDS and Cancer Research Abstracts
Virology and AIDS Abstracts
MEDLINE - Academic
DatabaseTitleList MEDLINE
AIDS and Cancer Research Abstracts
Database_xml – sequence: 1
  dbid: NPM
  name: PubMed
  url: https://proxy.k.utb.cz/login?url=http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed
  sourceTypes: Index Database
– sequence: 2
  dbid: EIF
  name: MEDLINE
  url: https://proxy.k.utb.cz/login?url=https://www.webofscience.com/wos/medline/basic-search
  sourceTypes: Index Database
DeliveryMethod fulltext_linktorsrc
Discipline Biology
EndPage 125
ExternalDocumentID 10445295
Genre Journal Article
GroupedDBID ---
--K
--M
-~X
.1-
.55
.FO
.GJ
.~1
0R~
123
1B1
1P~
1RT
1~.
1~5
3O-
4.4
457
4G.
53G
5VS
7-5
71M
8P~
9JM
AACTN
AAEDT
AAEDW
AAIKJ
AAKOC
AALCJ
AALRI
AAOAW
AAQFI
AAQXK
AATLK
AAXKI
AAXUO
ABGRD
ABGSF
ABJNI
ABMAC
ABUDA
ABXDB
ACDAQ
ACGFS
ACNCT
ACRLP
ADBBV
ADEZE
ADMUD
ADQTV
ADUVX
AEBSH
AEHWI
AEKER
AENEX
AEQOU
AEVXI
AFCTW
AFFNX
AFJKZ
AFKWA
AFRHN
AFTJW
AFXIZ
AGHFR
AGRDE
AGUBO
AGYEJ
AHHHB
AIEXJ
AIKHN
AITUG
AJOXV
AJUYK
AKRWK
ALMA_UNASSIGNED_HOLDINGS
AMFUW
AMRAJ
ASPBG
AVWKF
AXJTR
AZFZN
BKOJK
BLXMC
CAG
CGR
COF
CS3
CUY
CVF
DU5
EBS
ECM
EFJIC
EIF
EJD
EO8
EO9
EP2
EP3
F5P
FA8
FDB
FEDTE
FGOYB
FIRID
FNPLU
FYGXN
G-2
G-Q
GBLVA
HLW
HMK
HMO
HVGLF
HZ~
IHE
J1W
KOM
L7B
LX3
M29
M41
MO0
N9A
NPM
O-L
O9-
OAUVE
OHT
OZT
P-8
P-9
P2P
PC.
Q38
R2-
RIG
ROL
RPZ
SAE
SBG
SCC
SDF
SDG
SES
SEW
SPCBC
SSA
SSU
SSZ
T5K
UHS
WH7
WUQ
X7M
XOL
Z5R
ZCA
ZGI
~G-
7U9
H94
7X8
ID FETCH-LOGICAL-p238t-3437e1e7434544ff8b4dc155cec57f599b7739333e209b36151ad66ab1aca4003
ISSN 0040-8166
IngestDate Fri Oct 25 04:21:20 EDT 2024
Sun Sep 29 07:35:18 EDT 2024
Sat Sep 28 08:37:18 EDT 2024
IsPeerReviewed true
IsScholarly true
Issue 2
Language English
LinkModel OpenURL
MergedId FETCHMERGED-LOGICAL-p238t-3437e1e7434544ff8b4dc155cec57f599b7739333e209b36151ad66ab1aca4003
Notes ObjectType-Article-2
SourceType-Scholarly Journals-1
ObjectType-Feature-1
content type line 23
ObjectType-Article-1
ObjectType-Feature-2
PMID 10445295
PQID 17269988
PQPubID 23462
PageCount 9
ParticipantIDs proquest_miscellaneous_69957513
proquest_miscellaneous_17269988
pubmed_primary_10445295
PublicationCentury 1900
PublicationDate 1999-04-01
PublicationDateYYYYMMDD 1999-04-01
PublicationDate_xml – month: 04
  year: 1999
  text: 1999-04-01
  day: 01
PublicationDecade 1990
PublicationPlace Scotland
PublicationPlace_xml – name: Scotland
PublicationTitle Tissue & cell
PublicationTitleAlternate Tissue Cell
PublicationYear 1999
SSID ssj0007417
Score 1.6724658
Snippet Particulate antigens of the Hepatitis C virus (HCV) are reported for the first time by transmission electron microscopy in Pichia pastoris. The yeast was...
SourceID proquest
pubmed
SourceType Aggregation Database
Index Database
StartPage 117
SubjectTerms Gene Expression
Hepacivirus - physiology
Hepatitis C virus
Humans
Microscopy, Immunoelectron
Pichia - ultrastructure
Pichia pastoris
Viral Core Proteins - biosynthesis
Viral Core Proteins - genetics
Viral Envelope Proteins - biosynthesis
Viral Envelope Proteins - genetics
Virion - ultrastructure
Virus Assembly
Title Ultrastructural and immunocytochemical evidences of core-particle formation in the methylotrophic Pichia pastoris yeast when expressing HCV structural proteins (core-E1)
URI https://www.ncbi.nlm.nih.gov/pubmed/10445295
https://search.proquest.com/docview/17269988
https://search.proquest.com/docview/69957513
Volume 31
hasFullText 1
inHoldings 1
isFullTextHit
isPrint
link http://utb.summon.serialssolutions.com/2.0.0/link/0/eLvHCXMwnV3LbtQwFLWGIiQ2iDfl6QULUJQySZzXElVTBtQWCWXQ7CLbY7eVqiSaSYWGBZ_Af_CX3GvnSal4bKJRZuSxco-uT-xzzyXkJZqqGfWU0j6W5OiVK8KYub7UoZeshK-MgenRcTRfsA_LcDmZfB-oli5qsSe__rau5H-iCvcgrlgl-w-R7QaFG_AZ4gtXiDBc_yrGi_N6za0DrHHPMAcBWPBRym2NrbCsF4BqOodurIZ8rdyqGayvXWz1jthRGt7h63VZnaJq_gy10E7FjZnIxtliqx_ny6ntDWA0tMWJM9__7AymYbwfUHoD7NX83cxrNxwaGpyZcBvY4clBByKOx2Mj6e077HXER9u5K-Wcc-dTueHjjfIjSNvtjvj7fjfDOiB0IpgmQ7Opi2eZwwwdeAMk-oN069m6z2bl9mwJ9aVFAVgpRBK1hFibif6kzZbqyH37-GN-sDg8zLPZMrtGrvuQuDBj7n3rJUPAvuJWhYlzbE1AQ_ZmPPrVryuGtmS3ya3mfYO-tfG-QyaquEtu2A6k23vkxy8QogAhehlCtIMQLTUdQYh2EKJnBQUI0TGEqIUQbSFEDYQoQoj2EKIAITqYRgsh-qoB0Ov7ZHEwy_bnbtO9w62ABtZuwIJYeQoYKgsZ0zoRbCWBvUolw1iHaSpidGMMAuVPUxEgs-arKOLC45LDyhI8IDtFWahHhIpIChXF8VSziHEWcZ1KpVWYSMFS4Ke75EX7uHPIjghcXqjyYpMDPY_SNEmu_gV8jUePwS55aOOUV9bmJfemDFUJ4eM_jv6E3Oyx_JTswNNSz4Cr1uK5AdBPVPieuQ
link.rule.ids 315,783,787,27936,27937
linkProvider Elsevier
openUrl ctx_ver=Z39.88-2004&ctx_enc=info%3Aofi%2Fenc%3AUTF-8&rfr_id=info%3Asid%2Fsummon.serialssolutions.com&rft_val_fmt=info%3Aofi%2Ffmt%3Akev%3Amtx%3Ajournal&rft.genre=article&rft.atitle=Ultrastructural+and+immunocytochemical+evidences+of+core-particle+formation+in+the+methylotrophic+Pichia+pastoris+yeast+when+expressing+HCV+structural+proteins+%28core-E1%29&rft.jtitle=Tissue+%26+cell&rft.au=Falcon%2C+V&rft.au=Garcia%2C+C&rft.au=de+la+Rosa%2C+MC&rft.au=Menendez%2C+I&rft.date=1999-04-01&rft.issn=0040-8166&rft.volume=31&rft.issue=2&rft.spage=117&rft.epage=125&rft_id=info:doi/10.1054%2Ftice.1999.0032&rft.externalDBID=NO_FULL_TEXT
thumbnail_l http://covers-cdn.summon.serialssolutions.com/index.aspx?isbn=/lc.gif&issn=0040-8166&client=summon
thumbnail_m http://covers-cdn.summon.serialssolutions.com/index.aspx?isbn=/mc.gif&issn=0040-8166&client=summon
thumbnail_s http://covers-cdn.summon.serialssolutions.com/index.aspx?isbn=/sc.gif&issn=0040-8166&client=summon