PUBLICATION

Gene expression and functional analysis of Aha1a and Aha1b in stress response in zebrafish

Authors

Xiao, H., Wang, H., He, Q., Zhou, J., Du, S.

ID

ZDB-PUB-220714-11

Date

2022

Source

Comparative biochemistry and physiology. Part B, Biochemistry & molecular biology 262: 110777 (Journal)

Registered Authors

Du, Shao Jun (Jim), Zhou, Jianfeng

Keywords

Aha1(activator of Hsp90 ATPase), Heat shock protein, Heat shock response, Myosin chaperone

MeSH Terms

Adenosine Triphosphatases/metabolism
Animals
Embryo, Nonmammalian/metabolism*
Gene Expression
HSP90 Heat-Shock Proteins/metabolism
Heat-Shock Response
Molecular Chaperones/metabolism*
Zebrafish*/genetics
Zebrafish*/metabolism
Zebrafish Proteins/metabolism*

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PubMed

35830921 Full text @ Comp. Biochem. Physiol. B Biochem. Mol. Biol.

Abstract

Activator of heat shock protein 90 (hsp90) ATPase (Aha1) is a Hsp90 co-chaperone required for Hsp90 ATPase activation. Aha1 is essential for yeast survival and muscle development in C. elegans under elevated temperature and hsp90-deficeiency induced stress conditions. The roles of Aha1 in vertebrates are poorly understood. Here, we characterized the expression and function of Aha1 in zebrafish. We showed that zebrafish genome contains two aha1 genes, aha1a and aha1b, that show distinct patterns of expression during development. Under normal the physiological condition, aha1a is primarily expressed in skeletal muscle cells of zebrafish embryos, while aha1b is strongly expressed in the head region. aha1a and aha1b expression increased dramatically in response to heat shock induced stress. In addition, Aha1a-GFP fusion protein exhibited a dynamic translocation in muscle cells in response to heat shock. Moreover, upregulation of aha1 expression was also observed in hsp90a1 knockdown embryos that showed a muscle defect. Genetic studies demonstrated that knockout of aha1a, aha1b or both had no detectable effect on embryonic development, survival, and growth in zebrafish. The aha1a and aha1b mutant embryos showed normal muscle development and stress response in response to heat shock. Single or double aha1a and aha1b mutants could grow into normal reproductive adults with normal skeletal muscle structure and morphology compared with wild type control. Together, data from these studies indicate that Aha1a and Aha1b are involved in stress response. However, they are dispensable in zebrafish embryonic development, growth, and survival.

Genes / Markers

Marker	Marker Type	Name
ahsa1a	GENE	AHA1, activator of heat shock protein ATPase homolog 1a
ahsa1b	GENE	AHA1, activator of heat shock protein ATPase homolog 1b
dnaja	GENE	DnaJ heat shock protein family (Hsp40) member A
hsp90aa1.1	GENE	heat shock protein 90, alpha (cytosolic), class A member 1, tandem duplicate 1
hsp90aa1.2	GENE	heat shock protein 90, alpha (cytosolic), class A member 1, tandem duplicate 2
hsp90b1	GENE	heat shock protein 90, beta (grp94), member 1
hspa8b	GENE	heat shock protein family A (Hsp70) member 8b
myom3	GENE	myomesin 3

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Figures

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Expression

Gene	Fish	Conditions	Stage	Qualifier	Anatomy	Assay	Figure
ahsa1a	ahsa1a^mb23/mb23	standard conditions	Prim-5	Not Detected	skeletal muscle	ISH	Fig. 7
ahsa1a	ahsa1a^mb23/mb23	standard conditions	Prim-5		whole organism	RTPCR	Fig. 7
ahsa1a	ahsa1a^mb24/mb24	standard conditions	Prim-5		skeletal muscle	ISH	Fig. 7
ahsa1a	ahsa1a^mb24/mb24	standard conditions	Prim-5		whole organism	RTPCR	Fig. 7
ahsa1a	WT	standard conditions	Adult		brain	RTPCR	Fig. 2
ahsa1a	WT	standard conditions	Adult		eye	RTPCR	Fig. 2
ahsa1a	WT	standard conditions	Adult		gill	RTPCR	Fig. 2
ahsa1a	WT	standard conditions	Adult		gut	RTPCR	Fig. 2
ahsa1a	WT	standard conditions	Adult		heart	RTPCR	Fig. 2
ahsa1a	WT	standard conditions	Adult		integument	RTPCR	Fig. 2

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Phenotype

Fish	Conditions	Stage	Phenotype	Figure
WT	heat shock	Prim-5	skeletal muscle ahsa1a expression increased amount, abnormal	Fig. 3
WT	heat shock	Prim-5	skeletal muscle ahsa1a expression increased distribution, abnormal	Fig. 3
WT	heat shock	Prim-5	skeletal muscle hsp90aa1.1 expression increased amount, abnormal	Fig. 11
WT	heat shock	Prim-5	skeletal muscle hsp90aa1.1 expression increased distribution, abnormal	Fig. 11
WT	heat shock	Prim-5	whole organism ahsa1b expression increased amount, abnormal	Fig. 3
WT	heat shock	Prim-5	whole organism ahsa1b expression increased distribution, abnormal	Fig. 3
WT	heat shock	Prim-5	whole organism dnaja expression increased amount, abnormal	Fig. 11
WT	heat shock	Prim-5	whole organism dnaja expression increased distribution, abnormal	Fig. 11
WT	heat shock	Long-pec	head ahsa1b expression increased amount, abnormal	Fig. 3
WT	heat shock	Long-pec	head ahsa1b expression increased distribution, abnormal	Fig. 3

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Mutations / Transgenics

Allele	Construct	Type	Affected Genomic Region
mb21		Indel	ahsa1a
mb22		Small Deletion	ahsa1a
mb23		Insertion	ahsa1a
mb24		Small Deletion	ahsa1a
mb25		Small Deletion	ahsa1b
mb26		Indel	ahsa1b
mn0067Gt	Gt(GBT-rp2)	Transgenic Insertion	myom3

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Human Disease / Model

Sequence Targeting Reagents

Target	Reagent	Reagent Type
ahsa1a	CRISPR1-ahsa1a	CRISPR
ahsa1a	CRISPR2-ahsa1a	CRISPR
ahsa1a	TALEN1-ahsa1a	TALEN
ahsa1b	CRISPR1-ahsa1b	CRISPR
hsp90aa1.1	MO4-hsp90aa1.1	MRPHLNO
hsp90aa1.2	MO2-hsp90aa1.2	MRPHLNO

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Fish

Fish
ahsa1a^mb23/mb23
ahsa1a^mb24/mb24
ahsa1b^mb25/mb25
ahsa1b^mb25/mb25; ahsa1a^mb23/mb23
ahsa1b^mb26/mb26
WT
WT + MO2-hsp90aa1.2
WT + MO4-hsp90aa1.1

Antibodies

Orthology

Engineered Foreign Genes

Marker	Marker Type	Name
GFP	EFG	GFP
mRFP1	EFG	mRFP1

Mapping

Xiao et al., 2022 ZDB-PUB-220714-11 PMID:35830921

Gene expression and functional analysis of Aha1a and Aha1b in stress response in zebrafish

Xiao et al., 2022

ZDB-PUB-220714-11
PMID:35830921