TY - JOUR
T1 - Activation-induced cytidine deaminase targets DNA at sites of RNA polymerase II stalling by interaction with Spt5
AU - Pavri, Rushad
AU - Gazumyan, Anna
AU - Jankovic, Mila
AU - Di Virgilio, Michela
AU - Klein, Isaac
AU - Ansarah-Sobrinho, Camilo
AU - Resch, Wolfgang
AU - Yamane, Arito
AU - San-Martin, Bernardo Reina
AU - Barreto, Vasco
AU - Nieland, Thomas J.
AU - Root, David E.
AU - Casellas, Rafael
AU - Nussenzweig, Michel C.
N1 - Funding Information:
We thank members of the Nussenzweig lab for helpful discussions, Klara Velinzon for FACS sorting, and Tom Eisenreich and David Bosque for animal management. We thank Drs Jayanta Chaudhuri and Urszula Nowak for ChIP protocols and anti-AID antibody, Dr. Sohail Malik for generously providing purified recombinant DSIF and Dr. Alan Derr for assistance with informatics. M.D.V. is a fellow of the American-Italian Cancer Foundation. R.P was a recipient of The Irvington Institute Postdoctoral Fellowship of the Cancer Research Institute. The work was supported by NIH grant (AI037526) to M.C.N. M.C.N. is a Howard Hughes Medical Institute Investigator.
PY - 2010/10
Y1 - 2010/10
N2 - Activation-induced cytidine deaminase (AID) initiates antibody gene diversification by creating U:G mismatches. However, AID is not specific for antibody genes; Off-target lesions can activate oncogenes or cause chromosome translocations. Despite its importance in these transactions little is known about how AID finds its targets. We performed an shRNA screen to identify factors required for class switch recombination (CSR) of antibody loci. We found that Spt5, a factor associated with stalled RNA polymerase II (Pol II) and single stranded DNA (ssDNA), is required for CSR. Spt5 interacts with AID, it facilitates association between AID and Pol II, and AID recruitment to its Ig and non-. Ig targets. ChIP-seq experiments reveal that Spt5 colocalizes with AID and stalled Pol II. Further, Spt5 accumulation at sites of Pol II stalling is predictive of AID-induced mutation. We propose that AID is targeted to sites of Pol II stalling in part via its association with Spt5.
AB - Activation-induced cytidine deaminase (AID) initiates antibody gene diversification by creating U:G mismatches. However, AID is not specific for antibody genes; Off-target lesions can activate oncogenes or cause chromosome translocations. Despite its importance in these transactions little is known about how AID finds its targets. We performed an shRNA screen to identify factors required for class switch recombination (CSR) of antibody loci. We found that Spt5, a factor associated with stalled RNA polymerase II (Pol II) and single stranded DNA (ssDNA), is required for CSR. Spt5 interacts with AID, it facilitates association between AID and Pol II, and AID recruitment to its Ig and non-. Ig targets. ChIP-seq experiments reveal that Spt5 colocalizes with AID and stalled Pol II. Further, Spt5 accumulation at sites of Pol II stalling is predictive of AID-induced mutation. We propose that AID is targeted to sites of Pol II stalling in part via its association with Spt5.
KW - DNA
KW - Molimmuno
UR - http://www.scopus.com/inward/record.url?scp=77957239251&partnerID=8YFLogxK
U2 - 10.1016/j.cell.2010.09.017
DO - 10.1016/j.cell.2010.09.017
M3 - Article
C2 - 20887897
AN - SCOPUS:77957239251
SN - 0092-8674
VL - 143
SP - 122
EP - 133
JO - Cell
JF - Cell
IS - 1
ER -
