Changes in the proteome of Huh7 cells induced by transient expression of hepatitis D virus RNA and antigens.
Hepatitis delta virus (HDV) infection of human hepatocytes infected with the hepatitis B virus (HBV) is associated with increased liver damage and risk of fulminant disease. Although considerable progress has been made towards the elucidation of the mechanisms of HDV replication and pathogenesis, little is still known about the host factors involved in the different steps of the replication cycle. Here, we made use of a proteomic approach to analyse the global alterations in protein expression that arise in human hepatocytes separately transfected with each of the HDV components. Huh7 cells were transiently transfected with plasmids that code for the small delta antigen (S-HDAg), large delta antigen (L-HDAg), genomic RNA (gRNA), and antigenomic RNA (agRNA), respectively. Total protein extracts were separated by 2-DE and differentially expressed spots were identified by MALDI-TOF followed by database searching. We identified 32 proteins known to be involved in different pathways namely nucleic acid metabolism, protein metabolism, transport, signal transduction, apoptosis, and cell growth. Moreover, the down regulation of hnRNP D, HSP105, and triosephosphate isomerase was further confirmed by Real time PCR.
Unidade de Biologia Molecular, Instituto de Higiene e Medicina Tropical, Universidade Nova de Lisboa, Lisboa, Portugal.
Sérgio
S
Mota
Marta
M
Mendes
Deborah
D
Penque
Ana V
AV
Coelho
Celso
C
Cunha
D045744:Cell Line, Tumor; D015180:Electrophoresis, Gel, Two-Dimensional; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D003698:Hepatitis Delta Virus / Q000235:genetics* / Q000378:metabolism*; D035923:Hepatitis delta Antigens / Q000235:genetics / Q000378:metabolism*; D022781:Hepatocytes / Q000378:metabolism* / Q000821:virology; D006801:Humans; D013058:Mass Spectrometry; D010957:Plasmids / Q000235:genetics; D011506:Proteins / Q000302:isolation & purification; D020543:Proteome / Q000378:metabolism*; D012367:RNA, Viral / Q000235:genetics* / Q000378:metabolism; D015203:Reproducibility of Results; D014162:Transfection
D035923:Hepatitis delta Antigens; D011506:Proteins; D020543:Proteome; D012367:RNA, Viral