Characterisation of mutations in the CBP-gene in Rubinstein-Taybi syndrome and tumors.

Department                                  :  Dept. of Human and Clinical Genetics

Project supervisor                      :  Dr. D.J.M. Peters/ Dr. Jeroen H. Roelfsema

Address                                       :  Sylvius Laboratories, Wassenaarseweg 72, 2333 AL Leiden

Telephone number                     :  071-5276048

Fax number                                  :  071-5276075

E-mail address                             :


Background of research project     :

Rubinstein-Taybi syndrome (RTS) is characterised by mental retardation and specific skeletal malformations  Microdeletions, translocations and point mutations deleting or truncating one copy of CBP are responsible for RTS in 30% of the patients we have examined. However, a significant part of CBP has not yet been screened. A large number of binding domains for interacting proteins, have been mapped accurately. Mutations in one or more of these domains may account for a large number of as yet unidentified mutations. One of the characteristics of Rubinstein-Taybi syndrome is a predisposition for tumor development. This may be explained by mutation of both copies of the CBP gene in the tumors. We would like to know whether  CBP has a role in tumor development or progression in tunors in RTS patients and in sporadic tumors (keloids, breast cancer, cervix cancer).

Research questions               :

We hypothesize that missense mutations in functional domains of CBP, which have not yet been screened, can be responsible for RTS in a subset of patients. We further hypothesize that tumors in RTS patients can be caused by the loss of the unaffected copy of the CBP gene.


Project description and experimental design                   :

Intronic primers will be selected for the different exons of CBP.  For mutation analysis fragments generated via the  polymerase chain reaction (PCR) will be analysed by electrophoresis on agarose-gels and on polyacrylamide gels (Denaturing gradient gel electrophoresis, Single Strand Conformation Polymorphism analysis). Mutations will be analysed using an automatic sequencer. Whenever possible the mutation will be confirmed by digesting the DNA with a restriction enzyme.


Techniques to be applied              :

Polymerase Chain Reaction (PCR), Multiplex PCR, agarose gel electrophoresis, restriction enzyme digestion of DNA, Single stranded conformation polymorphism analyse (SSCP), Denaturing gradient gel electrophoresis. Analysis of sequence and fragment runs on ABI automated sequencers with specialized software.