Dual inhibitory effects of APOBEC family proteins on retrotransposition of mammalian endogenous retroviruses.
We demonstrated previously that the cytosine deaminase APOBEC3G inhibits retrotransposition of two active murine endogenous retroviruses, namely intracisternal A-particles (IAP) and MusD, in an ex vivo assay where retrotransposition was monitored by selection of neo-marked elements. Sequencing of the transposed copies further disclosed extensive editing, resulting in a high load of G-to-A mutations. Here, we asked whether this G-to-A editing was associated with an impact of APOBEC3G on viral cDNA yields. To this end, we used a specially designed quantitative PCR method to selectively measure the copy number of transposed retroelements, in the absence of G418 selection. We show that human APOBEC3G severely reduces the number of MusD and IAP transposed cDNA copies, with no effect on the level of the intermediate RNA transcripts. The magnitude of the decrease closely parallels that observed when transposed copies are assayed by selection of G418-resistant cells. Moreover, sequencing of transposed elements recovered by PCR without prior selection of the cells reveals high-level editing. Using this direct method with a series of cytosine deaminases, we further demonstrate a similar dual effect of African green monkey APOBE3G, human APOBEC3F and murine APOBEC3 on MusD retrotransposition, with a distinct extent and site specificity for each editing activity. Altogether the data demonstrate that cytosine deaminases have a protective effect against endogenous retroviruses both by reducing viral cDNA levels and by introducing mutations in the transposed copies, thus inactivating them for subsequent rounds of retrotransposition. This dual, two-step effect likely participates in the efficient defense of the cell genome against invading endogenous retroelements.
Unité des Rétrovirus Endogènes et Eléments Rétroïdes des Eucaryotes Supérieurs, CNRS UMR 8122 Institut, Gustave Roussy, 94805 Villejuif, France.
Cécile
C
Esnault
Jean
J
Millet
Olivier
O
Schwartz
Thierry
T
Heidmann
D000071480:APOBEC-3G Deaminase; D000818:Animals; D002522:Chlorocebus aethiops; D003564:Cytidine Deaminase / Q000378:metabolism; D043525:Cytosine Deaminase / Q000378:metabolism*; D020077:Endogenous Retroviruses / Q000235:genetics*; D006367:HeLa Cells; D006801:Humans; D051379:Mice; D009700:Nucleoside Deaminases / Q000378:metabolism*; D016133:Polymerase Chain Reaction / Q000379:methods; D017393:RNA Editing; D012097:Repressor Proteins / Q000378:metabolism*
D012097:Repressor Proteins; D009700:Nucleoside Deaminases; C492906:APOBEC3F protein, human; D043525:Cytosine Deaminase; D000071480:APOBEC-3G Deaminase; C464924:APOBEC3G protein, human; C503504:Apobec3 protein, mouse; D003564:Cytidine Deaminase