title: Calcium enhances mouse keratinocyte differentiation in vitro to differentially regulate expression of papillomavirus authentic and codon modified L1 genes.
abstract: Here, we first wished to establish for mouse primary keratinocytes (KCs) the Ca(2+) concentrations that were associated with KC differentiation in vitro. Using the range of Ca(2+) concentrations (0-6 mM) to differentiate primary KCs in culture to varying extents for 2 days, we then examined how KC differentiation impacted on expression of papillomavirus (PV) native (Nat) and codon modified (Mod) L1 genes. L1 mRNAs transcribed from either Nat or Mod L1 genes were present in similar amounts in KCs exposed to six Ca(2+) concentrations. However, expression of the L1 proteins from two Mod L1 genes were down-regulated, with no L1 signal detected in KCs exposed to 6 mM Ca(2+). In contrast, L1 proteins expressed from the two Nat L1 genes were not detectable in KCs without Ca(2+), but dramatically up-regulated as the KC cultures exposed to Ca(2+) from 0.5 to 2 mM, then down-regulated in KCs exposed to Ca(2+) from 4 to 6 mM. The different regulatory roles of the Ca(2+) in L1 protein expression from Nat and Mod L1 genes in cultured KCs were confirmed by TGF-beta1 experiments. We observed that aminoacyl-tRNAs (aa-tRNAs) from the 2 mM Ca(2+)-treated KCs only significantly enhanced the Nat L1 mRNAs translation in vitro, suggesting that aa-tRNAs play a differentially regulatory role in translations of the PV Nat and Mod L1 mRNAs. Importantly, the Ca(2+) experimental model provides evidence that mouse primary KCs could be transiently infected by BPV1 virus to express L1 mRNA and protein, which is very useful for future HPV virus infection study.
authors.affiliation: Diamantina Institute for Cancer, Immunology and Metabolic Medicine, The University of Queensland, Research Extension, Building 1, Princess Alexandra Hospital, Ipswich Road, Woolloongabba, Queensland 4102, Australia.
authors.forename: Ning-Xia
authors.initials: NX
authors.lastname: Fang
authors.forename: Wenyi
authors.initials: W
authors.lastname: Gu
authors.forename: Jianmin
authors.initials: J
authors.lastname: Ding
authors.forename: Nicholas A
authors.initials: NA
authors.lastname: Saunders
authors.forename: Ian H
authors.initials: IH
authors.lastname: Frazer
authors.forename: Kong-Nan
authors.initials: KN
authors.lastname: Zhao
mesh_terms: D000818:Animals; D002118:Calcium / Q000378:metabolism* / Q000494:pharmacology; D036022:Capsid Proteins / Q000235:genetics / Q000378:metabolism*; D002454:Cell Differentiation; D002478:Cells, Cultured; D003062:Codon; D015967:Gene Expression Regulation, Viral; D015603:Keratinocytes / Q000166:cytology / Q000821:virology*; D051379:Mice; D027383:Papillomaviridae / Q000235:genetics / Q000502:physiology*; D012333:RNA, Messenger / Q000378:metabolism
chemical_list: D036022:Capsid Proteins; D003062:Codon; C085217:L1 protein, Bovine papillomavirus; D012333:RNA, Messenger; D002118:Calcium