|
|
|
|
The DNA/RNA-dependent RNA polymerase QDE-1 generates aberrant RNA and dsRNA for RNAi in a process requiring replication protein a and a DNA helicase
|
|
|
|
|
|
|
|
نویسنده
|
liu y. ,lee h.-c. ,aalto a.p. ,yang q. ,chang s.-s. ,huang g. ,fisher d. ,cha j. ,poranen m.m. ,bamford d.h.
|
|
منبع
|
plos biology - 2010 - دوره : 8 - شماره : 10
|
|
چکیده
|
The production of aberrant rna (arna) is the initial step in several rnai pathways. how arna is produced and specifically recognized by rna-dependent rna polymerases (rdrps) to generate double-stranded rna (dsrna) is not clear. we previously showed that in the filamentous fungus neurospora,the rdrp qde-1 is required for rdna-specific arna production,suggesting that qde-1 may be important in arna synthesis. here we show that a recombinant qde-1 is both an rdrp and a dna-dependent rna polymerase (ddrp). its ddrp activity is much more robust than the rdrp activity and occurs on ssdna but not dsdna templates. we further show that replication protein a (rpa),a single-stranded dna-binding complex that interacts with qde-1,is essential for arna production and gene silencing. in vitro reconstitution assays demonstrate that qde-1 can produce dsrna from ssdna,a process that is strongly promoted by rpa. furthermore,the interaction between qde-1 and rpa requires the recq dna helicase qde-3,a homolog of the human werner/bloom syndrome proteins. together,these results suggest a novel small rna biogenesis pathway in neurospora and a new mechanism for the production of arna and dsrna in rnai pathways.© 2010 lee et al.
|
|
|
|
|
آدرس
|
department of physiology,university of texas southwestern medical center,dallas,tx, United States, department of physiology,university of texas southwestern medical center,dallas,tx,united states,university of massachusetts medical school,worcester,ma, United States, institute of biotechnology and department of biosciences,university of helsinki,helsinki, Finland, department of physiology,university of texas southwestern medical center,dallas,tx, United States, department of physiology,university of texas southwestern medical center,dallas,tx, United States, department of physiology,university of texas southwestern medical center,dallas,tx, United States, department of physiology,university of texas southwestern medical center,dallas,tx,united states,university of texas,austin,tx, United States, department of physiology,university of texas southwestern medical center,dallas,tx, United States, institute of biotechnology and department of biosciences,university of helsinki,helsinki, Finland, institute of biotechnology and department of biosciences,university of helsinki,helsinki, Finland
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
Authors
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|