SYNCRIP, a Member of the Heterogeneous Nuclear Ribonucleoprotein Family, Is Involved in Mouse Hepatitis Virus RNA Synthesis

Keum S. Choi; Akihiro Mizutani; Michael M. C. Lai

doi:10.1128/JVI.78.23.13153-13162.2004

FIG. 3.

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In vitro binding of the recombinant SYNCRIP and analysis of its binding sites on 5′-UTR. (A) Specific binding of recombinant F-SYN and N-SYN to 5′-UTR. Purified recombinant F-SYN and N-SYN (10 ng) were cross-linked to ³²P-labeled 5′-UTR (lanes 1 and 2) or vector pcDNA3.1 RNA transcript (200 nucleotides) (lanes 3 and 4). (B) Binding region of N-SYN in 5′-UTR. Purified N-SYN (10 ng) was cross-linked with 30 pmol of ³²P-labeled probes. R: UCUAA repeats. (C) Competition experiments. ³²P-labeled 5′-UTR (30 pmol) was UV cross-linked with 10 ng of purified N-SYN in the presence of unlabeled probes, 5′-UTR, c5′-UTR, 3′-UTR, c3′-UTR, and nonspecific RNA in serial dilutions (1-, 5-, 20-, and 50-fold). (-), UV cross-linked N-SYN without added unlabeled RNA.