Figure 1.

Schematic representation of gene constructs. (A) Rpp38 or portions of this protein, Rpp38(1-245) or Rpp38(246-283), was fused in-frame to GFP in pEGFP-C1 expression vector (see Materials and Methods). Numbers indicate positions of residues in the 283–amino acid Rpp38 polypeptide. NLS1 and NLS2 indicate putative NLSs. NS38 represents the amino acid sequence from position 260–283 of Rpp38. Substitution mutations of the nine lysines to asparagines in NS38 sequence, and their numbers, are presented in bold letters. Single substitution mutants of arginine (R13A), serine (S18A), threonine (T22A), and proline (P23A) to alanine are also shown. ATΔPP has substitution of arginine and lysine with alanine and threonine, respectively, and the two consecutive proline residues were deleted from NS38. (B) Full-length Rpp29 or the amino acids encompassing positions 52–85 or 63–85 of Rpp29 were fused in-frame with GFP in pEGFP-C1. The amino acid sequence of Rpp29 from position 63–85, designated NS29, is indicated. Several mutants of NS29 with single and multiple substitutions or deletions are shown. All gene fusion constructs were transcribed from a cytomegalovirus promoter-enhancer located upstream of GFP in the expression plasmid.