Molecular cloning and functional characterization of the human endogenous retrovirus K113.
The human endogenous retrovirus-K113 (HERV-K113) is the most complete HERV known to date. It contains open reading frames for all viral proteins. Depending on ethnicity, up to 30% of the human population carries the provirus on chromosome 19. To facilitate molecular and functional studies, we have cloned the HERV-K113 sequence into a small plasmid vector and characterized its functional properties. Here we show that based on a substantial LTR-promoter activity, full length messenger RNA and spliced env-, rec- and 1.5 kb (hel)-transcripts are produced. The envelope protein of HERV-K113 is synthesized as an 85 kDa precursor that is found partially processed. The accessory Rec protein is highly expressed and accumulates in the nucleus. Expression analysis revealed synthesis of the Gag precursor and the protease. However, the cloned HERV-K113 provirus is not replication competent. It carries inactivating mutations in the reverse transcriptase gene. These mutations can be reversed to reconstitute the active enzyme, but the reversion is not sufficient to reconstitute replication capacity of the virus.
Beimforde
Nadine
N
Robert Koch-Institut, ZBS 4, Nordufer 20, 13353 Berlin, Germany. BeimfordeN@rki.de
Hanke
Kirsten
K
Ammar
Ismahen
I
Kurth
Reinhard
R
Bannert
Norbert
N
D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism; D003001:Cloning, Molecular; D020077:Endogenous Retroviruses / Q000235:genetics* / Q000502:physiology*; D015870:Gene Expression; D015686:Gene Products, env / Q000378:metabolism*; D015683:Gene Products, gag / Q000737:chemistry / Q000378:metabolism*; D005814:Genes, Viral; D005822:Genetic Vectors; D006367:HeLa Cells; D006801:Humans; D007668:Kidney / Q000166:cytology; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010957:Plasmids; D011401:Promoter Regions, Genetic; D011533:Proviruses / Q000235:genetics; D022821:RNA Splice Sites; D012333:RNA, Messenger / Q000378:metabolism; D012367:RNA, Viral / Q000302:isolation & purification*; D012194:RNA-Directed DNA Polymerase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D020079:Terminal Repeat Sequences; D014158:Transcription, Genetic; D014162:Transfection; D014759:Viral Envelope Proteins / Q000378:metabolism*
C485839:ERVK-6 protein, human endogenous retrovirus; D015686:Gene Products, env; D015683:Gene Products, gag; D022821:RNA Splice Sites; D012333:RNA, Messenger; D012367:RNA, Viral; D014759:Viral Envelope Proteins; D012194:RNA-Directed DNA Polymerase