POINT SUMMARY JOURNAL Destabilization of linker histone H1.

2 is essential
for ATM activation and DNA damage repair

Introduction
1.Nucleosome ->> Basic unit chromatin ->> consist of octamer core histones and 146 bp
DNA.
There is many types of histones Ex. H1 , H2, H3, etc. but H1-H8 is core of nucleosome.
H1 ->> one of intra nucleosomal architecture protein ->> order accessibility and plasticity.
Has 11 Isoform. Has a main function in order genome stability and integrity.

2.Many evidence/proof show us H1 has participates in DNA repair

Decrease of H1 make high regulate homologous recombination machinery and enhance
resistance of DNA damage in yeast.
Decrease of H1 in ES mouse cell ensure DNA damage signaling and promote hyper
resistance of damaging DNA agents.
H1 also enhances the backup non-homologous end-joining (NHEJ) pathway by stimulating
the activities of DNA ligase IV and III.
3.Mechanism
Exact mechanism of main role action of H1 in DNA repair is still known limited.
One of H1 isoform subtype, H1.2 more prefer bind to AT DNA region which is more fragile
because lack of hydrogen bond. This is exhibit that H1.2 isoform have specific role in DNA
DAMAGE and repair.
4. Ataxia telangiectasia mutated (ATM)
a master kinase involved in the DNA damage response and repair, which exists as an
inactive homodimer or higher order multimer under basal/minimum conditions. Activation
of ATM is a complex and tightly regulated process that requires exposure of DNA breaks,
a cascade of acetylation and phosphorylation, and the assembly of the MRE11- RAD50-
NBS1 (MRN) complex. Numerous cellular processes have been implicated in ATM
activation and signaling, including PARP1-mediated poly-ADP-ribosylation (PARylation)
during DNA damage.
Note : MRN is complex of three proteins — Mre11, Rad50 and Nbs1 (also known as
Nibrin or p95). Mre11, Rad50 and Nbs1 are known tumor suppressors. Loss of function
of any of these proteins results in genome instability, the principal feature of cancer cells.
ATM activation may be associated with structural changes to chromatin as the induction
of perturbations to chromatin using sodium chloride (NaCl), chloroquine (CHQ) or histone
deacetylase (HDAC) inhibitors can potently activate ATM without eliciting DNA damage.
Chromatin interactions modulated by the nucleosome-binding protein HMGN1 through the
regulation of histone acetylation are also essential for ATM activation. Phosphorylation of
TIP60 by c-Abl upon chromatin disruption promotes ATM acetylation and subsequent
activation.
Basic assumption : H1 is critical for modulating chromatin dynamics and genome stability,
it is possible that H1, or one of its specific isoforms, may be associated with ATM
activation.
Discussion :
Evidence fact from experiment give results that H1.2 repressed activation and expression
of ATM. It is also protecting DNA packaging on chromatin level from strange ATM
treatment.
When DNA damage occurred, H1.2 will removed by it self and very PARylated. This
momentum make chromatin more accessible and let ATM activated.
This journal build up a new whole model to exhibit how H1.2 act as molecular brake on
ATM biding to MRN reaction.
ATM activation strongly associated with chromatin change packaging/conformation. But
from evidence and other study said other supporting factors also needed. For example of
some factor :
a. FoxO3a interacts with the FAT domain of ATM to regulate its dimerization and
subsequent activation
b. The E3 ligases Chfr and RNF8 synergistically control histone ubiquitination and
acetylation, leading to chromatin relaxation and ATM activation.
c. DNA PKcs directly phosphorylates ATM at multiple sites and inhibits ATM
activity.
Note : The DNA-dependent protein kinase (DNA–PK) is a nuclear serine/threonine protein
kinase that is activated upon association with DNA. Biochemical and genetic data have
revealed DNA–PK to be composed of a large catalytic subunit, termed DNA–PKcs, and a
regulatory factor termed Ku. In addition, recent work has implicated DNA–PK components
in a variety of other processes, including the modulation of chromatin structure and
telomere maintenance.
This journal exhibit us how ATM interact with H1.2(as a molecular brake) via specific and
specialized fragment in HEAT repeat domain. This protein act as enzymes like mediator
between H1.2 histones with ATM. The process involve regulate binding site of ATM as a
substrate. As a molecular brake, H1.2 compete with MRN to binding at ATM active site
(mediated by HEAT). The result of this process make separated or impaired ATM and
MRN and H1.2 as a inhibitor factor for ATM.
The existence of H1.2 as an inhibitory factor of ATM activation is supported by the theory
that the DNA damage response should be tightly checked and spatiotemporally regulated
to ensure optimized DNA repair.
Note : The HEAT repeat is a tandemly repeated, 37-47 amino acid long module occurring
in a number of cytoplasmic proteins, including the four name-giving proteins huntingtin,
elongation factor 3 (EF3), the 65 Kd alpha regulatory subunit of protein phosphatase 2A
(PP2A) and the yeast PI3-kinase TOR1. Arrays of HEAT repeats consists of 3 to 36 units
forming a rod-like helical structure and appear to function as protein-protein interaction
surfaces. It has been noted that many HEAT repeat-containing proteins are involved in
intracellular transport processes.
Strong antagonist interaction between ATM and H1.2 also show when ATM was activated.
On ATM artificially activated stage (usually use NaCL and HDAC), H1.2became
destabilization and lose the binding ability into chromatin . This fact prove us ATM
dependent DNA damage response can be activated without DNA break.
Interaction between MRN complex and ATM is absolutely required for ATM activation.
MRN complex is very sensitive to DNA damage and conformation. MRN was activated
ATM by positive feedback loop.
According to this journal, H1.2 regulate ATM via MRN activated dependent directly. This
is called main standard pathway. This pathway doesn’t disturb recruitment of MRN in other
location (still in same chromatin). These fact give us description about ATM activation
whereby use other pathway. The alternative ATM regulation pathway by H1.2 occurred
when ATM dependent MRN regulation pathway disturb by other molecule or in very
fluctuate MRN amount. For example
H1.2 will prefer inhibits H4K16 acetylation by high regulate expression of histone
deacetylase SIRT1 and HDAC1. H4K16 acetylation recently known other critical ATM
regulation support molecule.
Generally how H1 histones protein can contribute massive and maximally in DNA damage
and repair response is still known limited and elusive. However, the must important in
genome ability to face DNA damage, genome integrity must be maintenance carefully. In
this case H1.2 have important also elusive role to keep genome stability and integrity.
For example : Although H1.2 KO did not alter general chromatin structure or cell survival
under unstressed conditions, according this experiment results, together with previous
reports, show that deletion of H1.2 leads to cell cycle progression defects.
This suggests that H1.2 may keep critical functions in normall cells. Data derived from
Drosophila models showed that H1 counters genome instability through inhibition of R-
loop formation. This is in accord with our data that H1.2 potently inhibits ATM activation,
because R-loops are known to activate ATM. Hence it is possible that different H1 variants
may function distinctly at different stages of the DNA damage response and repair
Note : R loops are three-stranded nucleic acid structures that comprise nascent RNA
hybridized with the DNA template, leaving the nontemplate DNA single-stranded. R loops
form naturally during transcription even though their persistent formation can be a risky
outcome with deleterious effects on genome integrity. On the other hand, over the last few
years, an increasingly strong case has been built for R loops as potential regulators of gene
expression.
This complex mechanism will not occurred if there have not specific molecule reporter and
receiver However, in genome packaging, PARP1 do this function as first receiver DNA
damage signaling to other relay molecules. PARP1 has same function both in SSB or DSB
DNA.).