Analysis of the structural relationships between the DNA-binding phosphoproteins pp42, pp43 and pp44 by in situ peptide mapping.
A structural homology is established between three DNA-binding phosphoproteins located in the 42 to 44 kDa range, referred to as pp42, pp43 and pp44, from Chironomus tentans salivary gland cells by in situ peptide mapping. The staining patterns of pp42, pp43 and pp44 which resulted from digestion with Staphylococcus aureus V8, trypsin or papain proteases show the presence of 8 to 15 spots majority of which have identical mobility. In the patterns of the digests generated by treatments with trypsin about 10 spots appear in common between any pair of the protein substrates. In addition, each pattern includes two to three peptides of mobility not present in the other. Thus the peptide mapping of pp42, pp43 and pp44 based on the staining patterns of proteolytic digests suggest the existence of structural homology between the three unlabelled substrates. The proteolytic peptides carrying the rapidly turning over phosphate groups form markedly different electrophoretic patterns than the unlabelled peptides visualized by staining. Treatment of 32P-labelled pp42, pp43 and pp44 with V8 generates only one labelled fragment in the 30 kD range. The cleavage patterns of pp44 produced by chymotrypsin or papain contain seven to ten labelled fragments while those of pp42 and pp43 contain only two. The 32P-labelled tryptic peptides of pp42, pp43 and pp44 exhibit a ladder pattern for each substrate which probably arise by a consecutive removal of 25 to 35 amino acid residues from the primary digestion products pp29, pp29.5 and pp30 by cleavage of four to five putative interdomain regions. The possibility that these three structurally related phosphoproteins belong to the category of transcription factors is discussed.
Egyhazi
E
E
Department of Medical Cell Biology, Karolinska Institute, Stockholm, Sweden.
Stigare
J
J
Holst
M
M
Pigon
A
A
D000818:Animals; D002683:Chironomidae / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000737:chemistry*; D005786:Gene Expression Regulation; D015394:Molecular Structure; D010449:Peptide Mapping; D010750:Phosphoproteins / Q000737:chemistry*; D010766:Phosphorylation; D011499:Protein Processing, Post-Translational; D012471:Salivary Proteins and Peptides / Q000737:chemistry
D004268:DNA-Binding Proteins; D010750:Phosphoproteins; D012471:Salivary Proteins and Peptides