Covalent modifications of the core histone tails, includind acetylation, methylation, phosphorylation, ADP-ribosylation and ubiquitination govern the chromatine accessibility and create binding surfaces for protein recognition (such as the bromodomain far acetylated lysines and the chromodomain for methylated lysines), therefore dictating the rate of gene transcription. We investigated the impact of p210 bcr-abl tyrosine kinase of chronic myeloid leukemia (CMLl) on the acetylation and methylation at individual lysine residues within the NH2-terminal tails of histone H4, an integral component of transcriptional competence at the onset of S phase. To the purpose, we used RP-HPLC coupled with electrospray mass spectrometry (LCMS) to resolve the single H4 species (non-acetylated and mono-, di- or three-acetylated) and enzymatic digestion coupled with MALDI mass spectrometry to define the individuai lysine residues involved in acetylation and methylation processes. In single cell clones generated from the murine myeloid 32D cell line transducing a temperature-sensitive (ts) p210 bcr-abl construct (that owns constitutive tyrosine kinase activity at the permissive temperature of 33°C, but lacks it at the non- permissive temperature of 39°C) the inhibition of p210 bcr-abl tyrosine kinase induced by 1 M STI571 was associated with a significant (p
COVALENT MODIFICATIONS OF HISTONE H4 NH2-TERMINAL TAILS ASSOCIATED WITH P210 BCR-ABL TYROSINE KINASE
BRUSA, GIANLUCA;CALONGHI, NATALIA;SANTUCCI, MARIA ALESSANDRA
2004
Abstract
Covalent modifications of the core histone tails, includind acetylation, methylation, phosphorylation, ADP-ribosylation and ubiquitination govern the chromatine accessibility and create binding surfaces for protein recognition (such as the bromodomain far acetylated lysines and the chromodomain for methylated lysines), therefore dictating the rate of gene transcription. We investigated the impact of p210 bcr-abl tyrosine kinase of chronic myeloid leukemia (CMLl) on the acetylation and methylation at individual lysine residues within the NH2-terminal tails of histone H4, an integral component of transcriptional competence at the onset of S phase. To the purpose, we used RP-HPLC coupled with electrospray mass spectrometry (LCMS) to resolve the single H4 species (non-acetylated and mono-, di- or three-acetylated) and enzymatic digestion coupled with MALDI mass spectrometry to define the individuai lysine residues involved in acetylation and methylation processes. In single cell clones generated from the murine myeloid 32D cell line transducing a temperature-sensitive (ts) p210 bcr-abl construct (that owns constitutive tyrosine kinase activity at the permissive temperature of 33°C, but lacks it at the non- permissive temperature of 39°C) the inhibition of p210 bcr-abl tyrosine kinase induced by 1 M STI571 was associated with a significant (pI documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
