title: Effect of arginine methylation on the RNA recognition and cellular uptake of Tat-derived peptides.
abstract: Arginine (Arg) methylation is a common post-translational modification that regulates gene expression and viral infection. The HIV-1 Tat protein is an essential regulatory protein for HIV proliferation, and is methylated in the cell. The basic region (residues 47-57) of the Tat protein contains six Arg residues, and is responsible for two biological functions: RNA recognition and cellular uptake. In this study, we explore the effect of three different methylation states at each Arg residue in Tat-derived peptides on the two biological functions. The Tat-derived peptides were synthesized by solid phase peptide synthesis. TAR RNA binding of the peptides was assessed by electrophoresis mobility shift assays. The cellular uptake of the peptides into Jurkat cells was determined by flow cytometry. Our results showed that RNA recognition was affected by both methylation state and position. In particular, asymmetric dimethylation at position 53 decreased TAR RNA binding affinity significantly, but unexpectedly less so upon asymmetric dimethylation at position 52. The RNA binding affinity even slightly increased upon methylation at some of the flanking Arg residues. Upon Arg methylation, the cellular uptake of Tat-derived peptides mostly decreased. Interestingly, cellular uptake of Tat-derived peptides with a single asymmetrically dimethylated Arg residue was similar to the native all Arg peptide (at 120 μM). Based on our results, TAR RNA binding apparently required both guanidinium terminal NH groups on Arg53, whereas cellular uptake apparently required guanidinium terminal NH₂ groups instead. These results should provide insight into how nature uses arginine methylation to regulate different biological functions, and should be useful for the development of functional molecules with methylated arginines.
authors: Department of Chemistry, National Taiwan University, Taipei 10617, Taiwan.
authors: Jhe-Hao
authors: JH
authors: Li
authors: Department of Chemistry, National Taiwan University, Taipei 10617, Taiwan.
authors: Wen-Chieh
authors: WC
authors: Chiu
authors: Department of Chemistry, National Taiwan University, Taipei 10617, Taiwan.
authors: Yun-Chiao
authors: YC
authors: Yao
authors: Department of Chemistry, National Taiwan University, Taipei 10617, Taiwan. Electronic address: rpcheng@ntu.edu.tw.
authors: Richard P
authors: RP
authors: Cheng
mesh_terms: D001120:Arginine / Q000378:metabolism*; D005434:Flow Cytometry; D015696:Gene Products, tat / Q000737:chemistry* / Q000378:metabolism*; D006801:Humans; D019169:Jurkat Cells; D008745:Methylation; D008968:Molecular Conformation; D012367:RNA, Viral / Q000737:chemistry / Q000378:metabolism*
chemical_list: D015696:Gene Products, tat; D012367:RNA, Viral; D001120:Arginine
keywords: Arginine methylation; Cellular uptake; RNA recognition; Tat-derived peptide