Evidence for polyphosphate in phosphorylated nonhistone nuclear proteins.
Structural studies of eucaryotic nuclear proteins have revealed the presence of bound polymeric phosphates. 32P-labeled and nonlabeled nonhistone nuclear proteins (NHPs) were isolated from rat liver nuclei and subjected to various controlled hydrolytic conditions. The analysis of protease-trypsin limit peptides revealed the presence of six phosphorylated, homogeneous fragments with phosphate/amino acid molar ratios greater than unity, ranging from 1.3 to 79. Alkaline beta elimination of phosphoester bonds released polymeric phosphates with chain lengths from 2 to over 200, as determined by using two-dimensional chromatographic analysis. The identity of these labeled polymeric phosphates was established to be polyphosphate by a number of criteria, including chromatographic mobility, gravimetric precipitation to constant specific activity, generation of orthophosphate on hydrolysis, and the determination of the delta H of hydrolysis of phosphoanhydride bonds. The evidence suggests that, in addition to the phosphomonoesters of serine and threonine, multiple phosphoanhydride linkages can result in the formation of polyphosphorylated NHPs. Previous investigators have demonstrated that exogenous, free polyphosphate causes destabilization of chromatin and enhancement of transcription in vitro. Although the function of the polyphosphorylated NHPs is currently unknown, such findings have possible functional implications with regard to the postulated role of NHPs as positive modifiers of gene expression.
D000596:Amino Acids / Q000302:isolation & purification; D000818:Animals; D001464:Barium; D002467:Cell Nucleus / Q000378:metabolism; D055598:Chemical Phenomena; D002621:Chemistry; D002868:Chromosomal Proteins, Non-Histone / Q000032:analysis*; D006868:Hydrolysis; D008099:Liver / Q000032:analysis; D010750:Phosphoproteins / Q000032:analysis*; D011122:Polyphosphates / Q000302:isolation & purification*; D051381:Rats
D000596:Amino Acids; D002868:Chromosomal Proteins, Non-Histone; D010750:Phosphoproteins; D011122:Polyphosphates; D001464:Barium