Bioinformatics Identification of HSP70 in Chicken (Gallus gallus
domesticus)
Ronistra Ginting
1
and Mohammad Basyuni
2
1
Doctoral Program of Agricultural Sciences, Faculty of Agriculture, Universitas Sumatera Utara, Padang Bulan, Medan
20155, Indonesia
2
Department of Forestry, Faculty of Forestry, Universitas Sumatera Utara, Jl. Tri Dharma Ujung No. 1 Medan, North
Sumatera 20155, Indonesia
Keywords: Gallus gallus domesticus, Heat Shock Protein 70, NCBI, chicken productivity.
Abstract: Increasing the ambient temperature beyond the comfortable temperature zone range for chickens causes
stress heat in poultry, resulting in a decrease in chicken productivity. The current study assesses of the
bioinformatics on the National Center for Biotechnology Information (NCBI) database as well as expected
the physicochemical of Heat Shock Protein 70 (HSP70)in chicken (Gallus gallus domesticus). Several
parameters of physicochemical HSP70-mRNA in chicken (G. gallus domesticus) were varied among the
genes observed. There is eight HSP70 induced from chicken (G. gallus domesticus) deposited in NCBI. The
length of the genes was alternated with the genes ascertained. Several lines of coded protein were 273 to
652 amino acid. The mitochondria target peptide value diversified from 0.010 to 0.309, signifying that it is
expected to be a presence. The present result indicated the prominence of the variation and role of a physical
and chemical characteristic of the distinct amino acids in protein dehydration genes as extreme climate
stress in chicken. The NCBI online available overall suite of online resources can be accessed for biological
information the data, including the PubMed database and additional NCBI resources, focus on literature for
Identification HSP70 Gene in G. gallus domesticus results detected in 8 databases. The literature contained,
34 PubMed, 1 RefSeq transcripts, and 1 RefSeq proteins.
1 INTRODUCTION
Stress due to environmental heat is a major problem
that often occurs in modern poultry farming, stress
due to rising environmental temperatures affecting
chickens, especially in the finisher phase, causing
chicken deaths and economic losses for chicken
farming companies (Lara and Rostagno, 2014).
Quails are also like that, and it is challenging to
reduce excessive body heat due to the influence of
high environmental temperatures. Information about
how to control the response of living creatures to
stress control genes is very little. Consider this, the
research conducted on HSP70 protein in chickens is
fundamental (Xie et al., 2014). The primary
oligonucleotides used were unique primers for
chickens, namely primers with 960-bp from HSP70
amplified. The results show 98% homology with
stress proteins HSP70 in G. gallus and 99%
homology with Numida meleagris (Gaviol et al.,
2008).
In the breathing mechanism, water vapor brought
along with the bloodstream and released while
breathing is vital to reduce excessive body heat.
However, water that comes out of breathing during
prolonged stress can cause a decrease in blood
volume and a lack of body fluids or dehydration as a
result of changes in venous return and blood
circulation. Blood flow will be more dependent on
the upper respiratory tract of the body as a result of a
decrease in blood volume which ultimately can lead
to hyperthermia and lead to the death of chickens.
Damage at the cellular level can also occur due to
physiological changes due to stress due to the
temperature of this environment. During heat stress
due to an increase in environmental temperature,
there is a drastic increase in HSP70 levels in the
chicken's body. Changes that occur at the cellular
level during stress that occur due to high ambient
temperatures include; membrane liquidity increases,
cell viability decreases, cell death matches the
intensity of heat stress, cell type, cytoskeleton
46
Ginting, R. and Basyuni, M.
Bioinformatics Identification of HSP70 in Chicken (Gallus gallus domesticus).
DOI: 10.5220/0008505300460050
In Proceedings of the International Conference on Natural Resources and Technology (ICONART 2019), pages 46-50
ISBN: 978-989-758-404-6
Copyright
c
2019 by SCITEPRESS – Science and Technology Publications, Lda. All rights reserved
modification, cell cycle stage, membrane component
destabilization, stress protein codon formation in the
cell nucleus (Velichko et al., 2013).
Stress protein or HSP70 is essential and is
present in all cells of organisms or living things. In
particular, HSP has a significant role in the complex
assembly of multiprotein, cell cycle control and
signaling, folding/opening proteins, cell protection
against stress/apoptosis and protein
transport/sequencing into the correct sub-cellular
compartment. It is well known that HSP is involved
in antigen presentation with the role of
accompanying and transferring antigen peptides to
class I and class II molecules from the main
histocompatibility complex. Extracellular HSP can
also stimulate the best antigen-presenting cells of the
immune system which include macrophages and
dendritic cells. Based on its molecular weight, HSP
can be grouped into HSP10, HSP40, HSP60, HSP70,
HSP90, etc. (Srivastava et al., 2006; Mansilla et al.,
2012)
In chickens, NTPDase-8 (Nucleoside
Triphosphate Diphosphohydrolase 8) is the outer
surface of cell nucleotides with a large extracellular
domain (ECD) containing the transmembrane
domain (TMD) in N-and C-term. The nature of
NTPDases-8 is not susceptible to antigens which
cause membrane disturbances, in contrast to other
cell surfaces which are very vulnerable. cDNA from
NTPDase-8 chicken is dissolved, and the protein is
purified. The pure soluble ATP-ase activity of
chicken results in NTPDase-8 less than 15% of
membrane-bound full-length enzymes, the result of
ATP, ADP, and P (i) inactivation. Mutants are
truncated in the large extracellular domain (ECD)
anchored by NTPDase original TMDs from chickens
or TMD from NTPDases that are different from
NTPDase2 taken from humans. The difference is
shown as the timeline for nonlinear ATP hydrolysis
(Li et al., 2009).
Selection of various types of chickens for years
has been made to choose chickens with higher
productivity. The selected chicken is very sensitive
and sensitive to extreme environmental changes in
its development environment. The global increase in
temperature that occurs at this time will greatly
affect the productivity of the chicken. Maintenance
with an open cage system will make chickens
susceptible to changes in environmental temperature
from outside the enclosure which will cause
chickens to experience stress due to heat. The group
of proteins that play an important role in normal
cellular physiology and as cell protection against
various stressors including heat stress is Heat Shock
Protein (HSP). The levels of various HSP groups in
chickens increase in almost all tissues to respond to
stress due to high ambient temperatures. This
increased HSP level will protect cellular proteins to
avoid damage due to heat or high temperatures.
(Shanmugam et al., 2017)
2 MATERIALS AND METHOD
2.1 Materials
Eight data on HSP70 in chicken (G. gallus
domesticus) from the NCBI database. The NCBI
reference sequence from DNA and amino acids were
used in this study:
1. G. gallus heat shock protein 70 (HSP70) mRNA,
complete cds 1,920 bp linear mRNA Accession:
FJ217667.1 GI: 208964709
2. G. gallus heat shock protein 70 (HSP70) mRNA,
complete cds 2,371 bp linear mRNA Accession:
EU747335.1 GI: 190576827
3. G. gallus cell membrane ecto-ATPase mRNA,
complete cds 2,352 bp linear mRNA. Accession:
U74467.1 GI: 1732248
4. Chicken 78-kD glucose-regulated protein,
complete cds 2,389 bp linear mRNA. Accession:
M27260.1 GI: 211826
5. G. gallus ecto-ATP-diphosphohydrolase mRNA,
complete cds 1,482 bp linear mRNA Accession:
AF426405.1 GI: 16518969
6. G. gallus ecto-ATP-diphosphohydrolase mRNA,
complete cds 1,570 bp linear mRNA Accession:
AF041355.1 GI: 3320414
7. TPA_exp: G. gallus RBJ protein (RBJ) mRNA,
complete cds 822 bp linear mRNA. Accession:
BK001282.1 GI: 42475925
8. G. gallus cpsmb7 mRNA for proteasome subunit
Z, complete cds 935 bp linear mRNA Accession:
AB098728.1 GI: 30268673
2.2 Physical and chemical
characteristics of the HSP70 in
chicken (G. gallus domesticus)
Protparam online (web.expasy.org/protparam/) was
used to determine the structure, physicochemical
characteristics of HSP70 chicken (G. gallus
domesticus). Important factors are calculated for a
number of amino acid, molecular weight, a total
number of atoms, half-life period, theoretical
Bioinformatics Identification of HSP70 in Chicken (Gallus gallus domesticus)
47
isoelectric point values, extinction coefficients,
predictable half-life, high average, Grand average of
hydropathicity instability and aliphatic index.
(Basyuni et al., 2017).
Table 1: Physicochemical characteristic of the HSP70 in chicken (G. gallus domesticus)
Variant
X1
X2
X3
X4
X5
X6
X7
Length of genes/bp
1,920
2,371
2,352
2,389
1,482
1,570
822
Number of amino acids
634
634
495
652
493
493
273
Molecular weight
69814.95
69845.92
54533.71
72018.51
54063.69
54034.79
30778.32
Theoretical pI
5.50
5.59
7.86
5.12
5.78
5.95
8.88
Total number of atoms
9831
9830
7658
10194
7561
7568
4322
Extinction coefficient-ac
35090
35090
77530
29005
88530
88530
30285
Extinction coefficient-ar
34840
34840
76780
28880
87780
87780
29910
Half-life period
30 h
30 h
30 h
30 h
30 h
30 h
30 h
Instability index
30.97
31.05
34.01
32.95
27.63
27.47
37.49
Aliphatic index
81.21
79.53
86.73
86.26
87.65
89.23
77.51
Grand average of hydropathicity
-0.485
-0.522
-0.006
-0.495
0.067
0.088
-0.410
Extinction coefficient-ac = assuming all pairs of Cys residues form cystines
Extinction coefficient-ar = assuming all Cys residues are reduced
2.3. Possible peptide transfer and
subcellular localization of HSP70.
in chicken (Gallus gallus
domesticus)
Transit peptides can be predicted using the online
targetP1.1 server (www.cbs.dtu.dk/services/targetp/).
This position is in accordance with the expected
existence of Len, mitochondrial targeting peptide
(mTP), and Signal peptide of the secretory pathway
(SP). Online predictor of subcellular protein
localization, PSORT (psort.hgc.jp/form.html), is also
used to control the subcellular determination of
protein-induced dehydration. (Basyuni and Wati,
2018).
3 RESULTS AND DISCUSSION
3.1 Physicochemical characteristics of
the HSP70 in chicken (G. gallus
domesticus)
Several characters of physicochemical HSP70
induced by Chicken (G. gallus domesticus was
shown in Table 1. The genes length changes with
ascertained genes. Individual lines are coded of
protein from 273 to 652 amino acid. Important notes
are the theoretical isoelectric point values,
heterogeneity of relative molecular weights,
extinction coefficients, instability coefficients, total
atomic numbers, and general hydropathy rates along
the genes analyzed. Despite the availability of a lot
of climate pressure related to HSP70 in chickens (G.
gallus domesticus), quantitative gene expression
analysis of these genes is only recently attempted for
the identification of candidate genes/factors that are
contributing to high-temperature tolerance. With the
advent of the qPCR technique, it is easier to quantify
each gene and establish its relevance under the given
stress situations.
3.2 The promising of potential transit
peptide of HSP70 (G. gallus
domesticus)
Transfer in chickens (G. gallus domesticus) can be
seen in Table 2. Three reliability was determined:
Len, secretory pathway signal peptides,
Mitochondrial, Other target peptides, Loc, RC.
ICONART 2019 - International Conference on Natural Resources and Technology
48
Table 2: The promising of potential transit peptide of HSP70 (G. gallus domesticus)
Variant
Reliability
Len
Mitochondrial
target peptide
Signal peptide of
secretory pathway
Other
Loc
RC
X1
634
0.059
0.133
0.870
-
2
X2
634
0.059
0.133
0.870
-
2
X3
495
0.042
0.938
0.028
S
1
X4
652
0.063
0.783
0.248
S
3
X5
493
0.010
0.962
0.115
S
1
X6
493
0.010
0.962
0.115
S
1
X7
273
0.303
0.046
0.787
-
3
X8
277
0.309
0.053
0.611
-
4
It should be noted that the target value of the
mitochondrial target diversified from 0.010 to 0.309,
indicating that it is expected to be present. This data
shows that Heat Shock Protein 70 peptide (HSP70)
in Chicken (G. gallus domesticus) plays a vital role
in the mechanism of tolerance of extreme weather
such as high ambient temperatures (Wang et al.,
2013).
3.3 Subcellular localization of HSP70 in
chicken (G. gallus domesticus)
There are four (4) variants (X1, X2, X7, X8)
were located in the cytoplasm (cyto), cytoplasm
nucleus (cyto-nucl) and mitochondrial inner
membrane (mito). One variant (X4) only located in
the Golgi membrane (golg). Nearly all variants (X1,
X2, X3, X4, X5, X6, and X8) where the HSP70
Peptide is located in peroxisomal (pero) and
extracellular (Extr), except the X7 variant is not
detected. There are three (3) Variants (X1, X2, and
X8) were located in the nucleus (nucl). Only two (2)
variants (X1, X2) were located in the cytoskeleton
(cysk).
Within the plasma membrane (plas) and the
endoplasmic reticulum membrane (ER) there are 3
variants, namely variants X3, X5 and X6 contained
in the plasma membrane and variants X4, X5 and
X6 are in the endoplasmic reticulum membrane,
while in Lysosomes (lyso) there are 4 variants
namely X3, X4, X5, and X6. Based on the results of
this study, data obtained that HSP70 peptides in
almost all variants were found in extracellular (Extr)
and proximal (pero), except that the X7 variant was
not found in either while the organelles that have the
HSP70 peptide-type are the Golgi membranes (golg)
which are only possessed by the X4 variant (Ghosh
et al., 2014).
Table 3: Subcellular localization of HSP70 in chicken (G. gallus domesticus)
Variant
Cyto
cyto-nucl
nucl
pero
cysk
extr
Mito
plas
E.R.
lyso
Golg
X1
19
14
7
2
2
1
1
-
-
-
-
X2
18.5
14
6.5
2
2
2
1
-
-
-
-
X3
-
-
-
6
-
2
-
18
-
6
-
X4
-
-
-
2
-
4
-
-
23
2
1
X5
-
-
-
4
-
1
-
22
4
1
-
X6
-
-
-
4
-
1
-
23
3
1
-
X7
5
5.5
-
-
-
-
25
-
-
-
-
X8
11
9
3
4
-
3
11
-
-
-
-
Note: Nucl = nucleus; Cyto = cytoplasm; Cyto-nucl = cytoplasm nucleus, Pero = peroxisomal; Cysk = cytoskeleton; Extr
= extracellular; Mito = mitochondrial inner membrane; Plas = plasma membrane; E.R. (m) = endoplas reticulum;
Lyso = lysosome; gol = golgi membrane; - : not detected
Bioinformatics Identification of HSP70 in Chicken (Gallus gallus domesticus)
49
4 CONCLUSIONS
Eight data were obtained about Heat Shock Protein
70 (HSP70) in chickens (G. gallus domesticus) from
the NCBI database. The target value of the
mitochondrial target is diversified from 0.010 to
0.309. This shows that HSP70 in Chicken (G. gallus
domesticus) plays an essential role in tolerance
mechanisms of extreme weather such as high
ambient temperatures. HSP70 peptides in almost all
variants were found in extracellular (Extr) and
peroxisal (pero), except that the X7 variant was not
found in either while the organelles that have the
HSP70 peptide-type are the Golgi membranes (golg)
which are only possessed by the X4 variant.
REFERENCES
Basyuni, M., Wasilah, M., and Sumardi. 2017.
Bioinformatics study of the mangrove actin genes. J.
Phys. Conf. Ser. 801, 012013.
Basyuni M., and Wati, R., 2018. Bioinformatics analysis
of the predicted polyprenol reductase genes in higher
plants J. Phys.: Conf. Ser. 978 012050
Gaviol, H.C., Gasparino, E., Prioli, A.J., and Soares MA.,
2008. Genetic evaluation of the HSP70 protein in the
Japanese quail (Coturnix japonica). Genet. Mol. Res.
7(1), 133-139.
Ghosh, A., Davey, M., Chute, I.C., Griffiths, S.G., Lewis,
S., Chacko, S., Barnett, D., Crapoulet, N., Fournier, S.,
Joy, A. and Caissie, M.C., 2014. Rapid isolation of
extracellular vesicles from cell culture and biological
fluids using a synthetic peptide with specific affinity
for heat shock proteins. PloS one, 9(10), e110443.
Lara, L., and Rostagno, M., 2013. Impact of heat stress on
poultry production. Animals, 3(2), 356-369.
Li, C.S., Lee, Y. and Knowles, A.F., 2009. The stability of
chicken nucleoside triphosphate diphosphohydrolase 8
requires both of its transmembrane domains.
Biochemistry, 49(1), 134-146.
Mansilla, M.J., Montalban, X. and Espejo, C., 2012. Heat
shock protein 70: roles in multiple sclerosis.
Molecular Medicine, 18(6), 1018-1028.
Shanmugam M., Rajkumar, U., Vinoth, A. 2017. Heat
Shock Protein and Thermal Stress in Chicken. In
book: Heat Shock Proteins in Veterinary Medicine and
Sciences, 179-193.
Srivastava, S., Li, Z., Ko, K., Choudhury, P., Albaqumi,
M., Johnson, A.K., Yan, Y., Backer, J.M., Unutmaz,
D., Coetzee, W.A. and Skolnik, E.Y., 2006. Histidine
phosphorylation of the potassium channel KCa3. One
by nucleoside diphosphate kinase B is required for
activation of KCa3. 1 and CD4 T cells. Molecular
Cell, 24(5), 665-675.
Velichko, A.K., Markova, E.N., Petrova, N.V., Razin,
S.V. and Kantidze, O.L., 2013. Mechanisms of heat
shock response in mammals. Cellular and molecular
life sciences, 70(22), 4229-4241.
Wang, S.H., Cheng, C.Y., Tang, P.C., Chen, C.F., Chen,
H.H., Lee, Y.P. and Huang, S.Y., 2013. Differential
gene expressions in testes of L2 strain Taiwan country
chicken in response to acute heat stress.
Theriogenology, 79(2), 374-382.
Xie, J., Tang, L., Lu, L., Zhang, L., Xi, L., Liu, H.C.,
Odle, J. and Luo, X., 2014. Differential expression of
heat shock transcription factors and heat shock
proteins after acute and chronic heat stress in laying
chickens (Gallus gallus). PLoS One, 9(7), e102204.
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