DNAPK-RPA Interactions (literature search)

Summary     References     Sequence     Structure

 

Summary:

RPA residues

DNA-PK binding site

Specific Reference

RPA32 (Thr-21 & Ser-33 are phosphorylated)

DNA-PK§

Hurwitz (May ’97)

More RPA32 phosphorylation sites (T21, S23, S29, S33, S11/12/13, S8, S4)

DNA-PK§

Dixon (Sep. ’97)

RPA70

DNA-PKcs§

Pommier (Mar. ’99)

§ - Exact domains of interaction are yet to be determined.

 

References:

 

A. Information on specific interaction sites

 

  1. Anderson – Nov. 1993.

DNA-PK tends to phosphorylate a Ser/Thr residue that is followed by a Glutamine residue (S/T-Q).                                                                             (TOP)

 

  1. Wold et al. – Aug. 1996.

All phosphorylation by DNA-PK in vitro occurs on RPA32 Nterminal (1-33 aa).

RPA32 Serines 33, 52, 72, 174 fit the consensus sequence (S/T-Q) for DNA-PK.

(TOP)

 

  1. Hurwitz et al. – May 1997.

DNA-PK phosphorylates RPA32 at Thr-21 and Ser-33.

Other Serines in RPA32 (1-37) are not phosphorylated by DNA-PK.   (TOP)

 

  1. Dixon et al. – Sep. 1997.

UV radiation causes hyperphosphorylation of RPA32N at following sites:

Thr-21, Ser-23, Ser-29, Ser-33, one of the three Ser-11/12/13.

Other probable sites are Ser-8 and Ser-4.                                                         (TOP)

 

  1. Pommier et al. – Mar. 1999.

“DNA-PK consists of a catalytic subunit (DNA-PKcs) and the Ku autoantigen”.

DNA-PKcs can form a multiprotein complex with RPA in untreated cells.

DNA-PKcs complexes with Ku (not RPA) in camptothecin treated cells.

RPA70 and DNAPKcs domains are essential for RPA:DNA-PK complex. (TOP)

 

B. General interaction information

 

  1. Kelly at al. – Dec. 1994.
  2. Hurwitz et al. – Sep. 1996. – Talks of cdc2-cyclinA-RPA interaction.
  3. Kirchgessner et al. – Nov. 1996. – Talks of RPA hyperphosphorylation.
  4. Yen et al. – Sep. 1998.
  5. Lee et al. – Jan. 1999. – Mutation in the Zinc-finger domain in RPA70 hinders phosphorylation by DNA-PK.
  6. Fackelmayer et al. – Mar. 1999. – Talks of hyperphosphorylation during apoptosis.
  7. Dixon et al. – Mar. 2003.
  8. Lees-Miller et al. – Feb. 2004.                                                             (TOP)

 

 

C. DNA-PK Sequence

 

Source: P78527 – DNA-PKcs on NCBI protein sequence database [gi: 38258929].

 

D. DNA-PK Structure

 

The tertiary structure of DNA-PK is yet to be determined. Interested people can obtain the secondary structure using SABLE (secondary structure prediction server). The protein is too big to be shown here.

The tertiary structure of the RPA32 N-terminal domain is also yet to be determined. (TOP)