Age-dependent deamidation of H1° histones in chromatin of mammalian tissues

Lindner, Herbert; Sarg, Bettina; Grunicke, H.; Helliger, Wilfried

doi:10.1007/s004320050261

Cited by 46 publications

(28 citation statements)

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“…This technique is particularly suitable for separating posttranslationally modified proteins (e.g. acetylated core (45) and H1 histones (46), methylated and deamidated histones (35,(47)(48)(49), and phosphorylated H1 proteins (34, 50)). As shown in Fig.…”

Section: Resultsmentioning

confidence: 99%

Testis-specific Linker Histone H1t Is Multiply Phosphorylated during Spermatogenesis

Sarg

Chwatal

Talasz

et al. 2009

Journal of Biological Chemistry

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During normal spermatogenesis, the testis-specific linker histone H1t appears at pachytene stage becomes phosphorylated in early spermatids and disappears in late spermatids. Using reversed-phase and hydrophilic interaction liquid chromatography, H1t from rat and mouse testes was isolated, subjected to enzymatic digestion, and analyzed by mass spectrometry. We observed different phosphorylated states of H1t (mono-, di-, and triphosphorylated) as well as the unphosphorylated protein.Tandem mass spectrometry and immobilized metal ion affinity chromatography experiments with MS/MS/MS and multistage activation were utilized to identify five phosphorylation sites on H1t from rats. Phosphorylation occurs on both serine and threonine residues, whereas only two of these sites were located on peptides containing the CDK consensus motif (S/T)PXZ. Rat H1t phosphorylation starts first by phosphorylation of the nonconsensus motif SPKS in the COOH-terminal domain, namely at Ser-140 and to a smaller degree at a further nonconsensus motif at Ser-186. This is followed by phosphorylation of Ser-177 and Thr-155, both located in CDK consensus motifs. A single phosphorylation site at Ser-8 in the NH 2 -terminal tail was also found. Mouse H1t lacks Ser-186 and is phosphorylated at up to four sites. In contrast to somatic linker histones, no strict order of increasing phosphorylation could be detected in H1t. Thus, it appears that not the order of up-phosphorylation but the number of the phosphate groups is necessary for regulated chromatin decondensation, thus facilitating the substitution of H1t by transition proteins and protamines.Mammalian spermatogenesis is a complex, strictly organized process, including proliferation and differentiation. After several mitotic divisions of spermatogonia, germ cells differentiate into spermatocytes and pass through two consecutive meiotic divisions, resulting in haploid round cells termed spermatids. These spermatids then evolve into highly condensed and transcriptionally inert sperm through a process known as spermiogenesis (for a review, see Ref.

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Section: Resultsmentioning

confidence: 99%

Testis-specific Linker Histone H1t Is Multiply Phosphorylated during Spermatogenesis

Sarg

Chwatal

Talasz

et al. 2009

Journal of Biological Chemistry

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“…In such cells, histones are thought to remodel chromatin to suppress transcription. Histone H1°ex-ists in liver and brain in two to four subfractions; the relative proportion of each varies with age (35,36). These subfractions are made up of native histone H1°, histone H1°that is acetylated at the N terminus, and histone H1°that is deamidated at Asn 3 and either acetylated or unacetylated at the N terminus (35,36).…”

Section: Asparagine Deamidation May Serve As Timer Of Chromatin Remodmentioning

confidence: 99%

“…Histone H1°ex-ists in liver and brain in two to four subfractions; the relative proportion of each varies with age (35,36). These subfractions are made up of native histone H1°, histone H1°that is acetylated at the N terminus, and histone H1°that is deamidated at Asn 3 and either acetylated or unacetylated at the N terminus (35,36). The abundance of the deamidated forms of histone H1°is 1.6-to 4-fold as high in the livers and 8-to 12-fold as high in the brains of 10-month-old rodents, respectively, compared with those of 20-day-old rodents (36).…”

Section: Asparagine Deamidation May Serve As Timer Of Chromatin Remodmentioning

confidence: 99%

“…These subfractions are made up of native histone H1°, histone H1°that is acetylated at the N terminus, and histone H1°that is deamidated at Asn 3 and either acetylated or unacetylated at the N terminus (35,36). The abundance of the deamidated forms of histone H1°is 1.6-to 4-fold as high in the livers and 8-to 12-fold as high in the brains of 10-month-old rodents, respectively, compared with those of 20-day-old rodents (36). The authors speculated that increased abundance of the deamidated forms in the older tissue might have a functional role in the chromatin remodeling that suppresses transcription in aging tissue.…”

Section: Asparagine Deamidation May Serve As Timer Of Chromatin Remodmentioning

confidence: 99%

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Chronoregulation by Asparagine Deamidation

Weintraub

Deverman

2007

Sci. STKE

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Every asparagine in every protein undergoes nonenzymatic deamidation to aspartate or isoaspartate at a rate determined by the surrounding protein structure and cellular environment. Under physiologic conditions, the deamidation half-life of individual asparagines in proteins is proposed to range from less than a day to several centuries. More than 200 proteins have been shown to undergo deamidation to a meaningful degree, and modeling predicts that hundreds more undergo deamidation at rates that have the potential to be of biological consequence. Because deamidation converts an asparagine into an aspartate or isoaspartate, it introduces a negative charge into a protein and results in the isomerization of a residue. Therefore, deamidation has the potential to change protein function. Additionally, deamidation is thought to render some proteins more susceptible to degradation. In most instances in which asparagine deamidation has been identified in vivo, it is involved in pathology. Hence, deamidation has been viewed primarily as a form of protein damage. However, the pervasiveness and evolutionary persistence of these unstable asparagines suggest that they may have a beneficial role. Notably, the change of even a single neighboring amino acid can have a marked effect on the rate of deamidation of an asparagine. Therefore, the underlying rate of deamidation of any asparagine is genetically programmable. This characteristic, combined with the wide range of deamidation rates that can be programmed, imparts to asparagines the potential to serve as molecular timers that regulate protein function and stability.

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“…With a hydrophobic counterion like the PFPA − anion, the migration order is analogous to RP-HPLC within the same side-chain type, whereas, with a hydrophilic counterion like the H 2 PO 4 − anion, the migration order is analogous to HILIC. The closest chromatography analogy to II-CZE is mixed-mode hydrophilic interaction-cation exchange chromatography (HILIC-CEX) [28][29][30][31][32][33][34]. In this method, the hydrophilic stationary phase has covalently attached sulfonate anions (SO 3 − ).…”

Section: Separative Mechanism In Ii-czementioning

confidence: 99%

Ion‐interaction CZE: The presence of high concentrations of ion‐pairing reagents demonstrates the complex mechanisms involved in peptide separations

2007

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We have furthered our understanding of the separative mechanism of a novel CE approach, termed ion-interaction CZE (II-CZE), developed in our laboratory for the resolution of mixtures of cationic peptides. Thus, II-CZE and RP-HPLC were applied to the separation of peptides differing by a single amino acid substitution in 10-and 12-residue synthetic model peptide sequences. Substitutions differed by a wide range of properties or side-chain type (e.g., alkyl side-chains, polar side-chains, etc.) at the substitution site. When carried out in high concentrations (400 mM) of pentafluoropropionic acid (PFPA), II-CZE separated peptides in order of increasing hydrophobicity when the substituted side-chains were of a similar type; when II-CZE was applied to the mixtures of peptides with substitutions of side-chains that differed in the type of functional group, there was no longer a correlation of electrophoretic mobility in II-CZE with relative peptide hydrophobicity, suggesting that a third factor is involved in the separative mechanism beyond charge and hydrophobicity. Interestingly, the hydrophobic PFPA − anion is best for separating peptides that differ in hydrophobicity with hydrophobic side-chains but high concentrations of the hydrophilic H 2 PO 4 − anion are best when separating peptides that differ in polar side-chains relative to hydrophobic side-chains. We speculate that differential hydration/dehydration properties of various side-chains in the peptide and the hydration/dehydration properties of the hydrophilic/hydrophobic anions as well as the electrostatic attractions between the peptide and the anions in solution all play a critical role in these solution-based effects.

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Age-dependent deamidation of H1° histones in chromatin of mammalian tissues

Cited by 46 publications

References 16 publications

Testis-specific Linker Histone H1t Is Multiply Phosphorylated during Spermatogenesis

Testis-specific Linker Histone H1t Is Multiply Phosphorylated during Spermatogenesis

Chronoregulation by Asparagine Deamidation

Ion‐interaction CZE: The presence of high concentrations of ion‐pairing reagents demonstrates the complex mechanisms involved in peptide separations

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