The RNA-dependent RNA polymerase essential for post-transcriptional gene silencing in Neurospora crassa interacts with replication protein A.
Post-transcriptional gene silencing (PTGS) pathways play a role in genome defence and have been extensively studied, yet how repetitive elements in the genome are identified is still unclear. It has been suggested that they may produce aberrant transcripts (aRNA) that are converted by an RNA-dependent RNA polymerase (RdRP) into double-stranded RNA (dsRNA), the essential intermediate of PTGS. However, how RdRP enzymes recognize aberrant transcripts remains a key question. Here we show that in Neurospora crassa the RdRP QDE-1 interacts with Replication Protein A (RPA), part of the DNA replication machinery. We show that both QDE-1 and RPA are nuclear proteins and that QDE-1 is specifically recruited onto the repetitive transgenic loci. We speculate that this localization of QDE-1 could allow the in situ production of dsRNA using transgenic nascent transcripts as templates, as in other systems. Supporting a link between the two proteins, we found that the accumulation of short interfering RNAs (siRNAs), the hallmark of silencing, is dependent on an ongoing DNA synthesis. The interaction between QDE-1 and RPA is important since it should guide further studies aimed at understanding the specificity of the RdRP and it provides for the first time a potential link between a PTGS component and the DNA replication machinery.
Dipartimento di Biotecnologie Cellulari ed Ematologia, Università La Sapienza, Rome, Italy.
Tony
T
Nolan
Germano
G
Cecere
Carmine
C
Mancone
Tonino
T
Alonzi
Marco
M
Tripodi
Caterina
C
Catalanotto
Carlo
C
Cogoni
D004271:DNA, Fungal / Q000096:biosynthesis / Q000737:chemistry; D005656:Fungal Proteins / Q000378:metabolism*; D015966:Gene Expression Regulation, Fungal; D020043:Genes, Essential; D009492:Neurospora crassa / Q000201:enzymology / Q000235:genetics*; D009687:Nuclear Proteins / Q000378:metabolism; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000378:metabolism; D012324:RNA-Dependent RNA Polymerase / Q000378:metabolism*; D012091:Repetitive Sequences, Nucleic Acid; D051716:Replication Protein A / Q000235:genetics / Q000378:metabolism*; D019076:Transgenes
D004271:DNA, Fungal; D005656:Fungal Proteins; D009687:Nuclear Proteins; D034741:RNA, Small Interfering; D051716:Replication Protein A; C119303:QDE1 protein, Neurospora crassa; D012324:RNA-Dependent RNA Polymerase