Bioinformatics Identification of HSP70 in Chicken (Gallus gallus

domesticus)

Ronistra Ginting

and Mohammad Basyuni

Doctoral Program of Agricultural Sciences, Faculty of Agriculture, Universitas Sumatera Utara, Padang Bulan, Medan

20155, Indonesia

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

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

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)

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

Length of genes/bp

1,920

2,371

2,352

2,389

1,482

1,570

822

935

Number of amino acids

634

495

652

493

273

277

Molecular weight

69814.95

69845.92

54533.71

72018.51

54063.69

54034.79

30778.32

29876.03

Theoretical pI

5.50

5.59

7.86

5.12

5.78

5.95

8.88

5.99

Total number of atoms

9831

9830

7658

10194

7561

7568

4322

4184

Extinction coefficient-ac

35090

77530

29005

88530

30285

18255

Extinction coefficient-ar

34840

76780

28880

87780

29910

17880

Half-life period

30 h

Instability index

30.97

31.05

34.01

32.95

27.63

27.47

37.49

20.31

Aliphatic index

81.21

79.53

86.73

86.26

87.65

89.23

77.51

86.64

Grand average of hydropathicity

-0.485

-0.522

-0.006

-0.495

0.067

0.088

-0.410

-0.121

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

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

634

0.059

0.133

0.870

634

0.059

0.133

0.870

495

0.042

0.938

0.028

652

0.063

0.783

0.248

493

0.010

0.962

0.115

493

0.010

0.962

0.115

273

0.303

0.046

0.787

277

0.309

0.053

0.611

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

18.5

6.5

5.5

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)

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.

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ICONART 2019 - International Conference on Natural Resources and Technology