Neospora caninum: Tachyzoites express a potent type-I nucleoside triphosphate hydrolase, but lack nucleoside diphosphate hydrolase activity

Takashi Asai; Daniel K. Howe; Kyoko Nakajima; Tomoyoshi Nozaki; Tsutomu Takeuchi; L. David Sibley

doi:10.1006/expr.1998.4346

Neospora caninum: Tachyzoites express a potent type-I nucleoside triphosphate hydrolase, but lack nucleoside diphosphate hydrolase activity

Takashi Asai, Daniel K. Howe, Kyoko Nakajima, Tomoyoshi Nozaki, Tsutomu Takeuchi, L. David Sibley

Research output: Contribution to journal › Article › peer-review

43 Scopus citations

Abstract

We have identified type I nucleoside triphosphate hydrolase (NTPase; EC 3.6.1.3) activity, previously thought to be restricted to the virulent strains of Toxoplasma gondii, in the cell extracts of Neospora caninum tachyzoites. Sequence analysis of a complete cDNA from Nc-1 strain indicated that N. caninum NTPases shared approximately 69% identity to the NTPases of T. gondii and are most similar to the NTPase-I isozyme. Southern blot analysis of genomic DNA and sequence analysis of two independent NTP clones from the Nc-1 strain revealed the presence of multiple genes, at least two of which are transcribed. Substrate specificity and K(m) values for MgATP² and MgADP^- hydrolysis for recombinant or partially purified native NcNTPase were the same as those for the type I isozyme (NTPase-I). Significantly, no type II enzyme (NTPase-II) activity for NDP hydrolysis was detected in cell extracts of N. caninum, although it is universally present in all T. gondii strains that have been tested. This intriguing difference between these two closely related apicomplexan parasites may provide insight into the function of the NTPases during intracellular parasitism.

Original language	English
Pages (from-to)	277-285
Number of pages	9
Journal	Experimental Parasitology
Volume	90
Issue number	3
DOIs	https://doi.org/10.1006/expr.1998.4346
State	Published - Nov 1998

Bibliographical note

Funding Information:
We thank Marilyn Levy for expert technical assistance with electron microscopy and Dr. Helen Profous-Juchelka, Merck Research Labs, for providing extracts of E. tenella. This work was supported in part by funds from the USDA (97-35204-4770), a National Grant-in-Aid from the Ministry of Education of Japan (10670236), the Japan Society for the Promotion of Science (JSPS-RFTF97L00701), and the Promotion of AIDS Research from The Ministry of Health and Welfare of Japan.

Funding

We thank Marilyn Levy for expert technical assistance with electron microscopy and Dr. Helen Profous-Juchelka, Merck Research Labs, for providing extracts of E. tenella. This work was supported in part by funds from the USDA (97-35204-4770), a National Grant-in-Aid from the Ministry of Education of Japan (10670236), the Japan Society for the Promotion of Science (JSPS-RFTF97L00701), and the Promotion of AIDS Research from The Ministry of Health and Welfare of Japan.

Funders	Funder number
Ministry of Education, Culture, Sports, Science & Technology, Japan	10670236
National Institute of Allergy and Infectious Diseases	R01AI034036
U.S. Department of Agriculture	97-35204-4770
Japan Society for the Promotion of Science	JSPS-RFTF97L00701
Ministry of Health, Labour and Welfare

Keywords

ATPase
Apicomplexan protozoa
Neosporosis
Purine salvage
Toxoplasma gondii

ASJC Scopus subject areas

Parasitology
Immunology
Infectious Diseases

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

Access to Document

10.1006/expr.1998.4346

Cite this

@article{3bb96cf934cb48cd8ccaba85a936fdf8,

title = "Neospora caninum: Tachyzoites express a potent type-I nucleoside triphosphate hydrolase, but lack nucleoside diphosphate hydrolase activity",

abstract = "We have identified type I nucleoside triphosphate hydrolase (NTPase; EC 3.6.1.3) activity, previously thought to be restricted to the virulent strains of Toxoplasma gondii, in the cell extracts of Neospora caninum tachyzoites. Sequence analysis of a complete cDNA from Nc-1 strain indicated that N. caninum NTPases shared approximately 69% identity to the NTPases of T. gondii and are most similar to the NTPase-I isozyme. Southern blot analysis of genomic DNA and sequence analysis of two independent NTP clones from the Nc-1 strain revealed the presence of multiple genes, at least two of which are transcribed. Substrate specificity and K(m) values for MgATP2 and MgADP- hydrolysis for recombinant or partially purified native NcNTPase were the same as those for the type I isozyme (NTPase-I). Significantly, no type II enzyme (NTPase-II) activity for NDP hydrolysis was detected in cell extracts of N. caninum, although it is universally present in all T. gondii strains that have been tested. This intriguing difference between these two closely related apicomplexan parasites may provide insight into the function of the NTPases during intracellular parasitism.",

keywords = "ATPase, Apicomplexan protozoa, Neosporosis, Purine salvage, Toxoplasma gondii",

author = "Takashi Asai and Howe, {Daniel K.} and Kyoko Nakajima and Tomoyoshi Nozaki and Tsutomu Takeuchi and Sibley, {L. David}",

note = "Funding Information: We thank Marilyn Levy for expert technical assistance with electron microscopy and Dr. Helen Profous-Juchelka, Merck Research Labs, for providing extracts of E. tenella. This work was supported in part by funds from the USDA (97-35204-4770), a National Grant-in-Aid from the Ministry of Education of Japan (10670236), the Japan Society for the Promotion of Science (JSPS-RFTF97L00701), and the Promotion of AIDS Research from The Ministry of Health and Welfare of Japan.",

year = "1998",

month = nov,

doi = "10.1006/expr.1998.4346",

language = "English",

volume = "90",

pages = "277--285",

number = "3",

}

TY - JOUR

T1 - Neospora caninum

T2 - Tachyzoites express a potent type-I nucleoside triphosphate hydrolase, but lack nucleoside diphosphate hydrolase activity

AU - Asai, Takashi

AU - Howe, Daniel K.

AU - Nakajima, Kyoko

AU - Nozaki, Tomoyoshi

AU - Takeuchi, Tsutomu

AU - Sibley, L. David

N1 - Funding Information: We thank Marilyn Levy for expert technical assistance with electron microscopy and Dr. Helen Profous-Juchelka, Merck Research Labs, for providing extracts of E. tenella. This work was supported in part by funds from the USDA (97-35204-4770), a National Grant-in-Aid from the Ministry of Education of Japan (10670236), the Japan Society for the Promotion of Science (JSPS-RFTF97L00701), and the Promotion of AIDS Research from The Ministry of Health and Welfare of Japan.

PY - 1998/11

Y1 - 1998/11

N2 - We have identified type I nucleoside triphosphate hydrolase (NTPase; EC 3.6.1.3) activity, previously thought to be restricted to the virulent strains of Toxoplasma gondii, in the cell extracts of Neospora caninum tachyzoites. Sequence analysis of a complete cDNA from Nc-1 strain indicated that N. caninum NTPases shared approximately 69% identity to the NTPases of T. gondii and are most similar to the NTPase-I isozyme. Southern blot analysis of genomic DNA and sequence analysis of two independent NTP clones from the Nc-1 strain revealed the presence of multiple genes, at least two of which are transcribed. Substrate specificity and K(m) values for MgATP2 and MgADP- hydrolysis for recombinant or partially purified native NcNTPase were the same as those for the type I isozyme (NTPase-I). Significantly, no type II enzyme (NTPase-II) activity for NDP hydrolysis was detected in cell extracts of N. caninum, although it is universally present in all T. gondii strains that have been tested. This intriguing difference between these two closely related apicomplexan parasites may provide insight into the function of the NTPases during intracellular parasitism.

AB - We have identified type I nucleoside triphosphate hydrolase (NTPase; EC 3.6.1.3) activity, previously thought to be restricted to the virulent strains of Toxoplasma gondii, in the cell extracts of Neospora caninum tachyzoites. Sequence analysis of a complete cDNA from Nc-1 strain indicated that N. caninum NTPases shared approximately 69% identity to the NTPases of T. gondii and are most similar to the NTPase-I isozyme. Southern blot analysis of genomic DNA and sequence analysis of two independent NTP clones from the Nc-1 strain revealed the presence of multiple genes, at least two of which are transcribed. Substrate specificity and K(m) values for MgATP2 and MgADP- hydrolysis for recombinant or partially purified native NcNTPase were the same as those for the type I isozyme (NTPase-I). Significantly, no type II enzyme (NTPase-II) activity for NDP hydrolysis was detected in cell extracts of N. caninum, although it is universally present in all T. gondii strains that have been tested. This intriguing difference between these two closely related apicomplexan parasites may provide insight into the function of the NTPases during intracellular parasitism.

KW - ATPase

KW - Apicomplexan protozoa

KW - Neosporosis

KW - Purine salvage

KW - Toxoplasma gondii

UR - http://www.scopus.com/inward/record.url?scp=0032212406&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0032212406&partnerID=8YFLogxK

U2 - 10.1006/expr.1998.4346

DO - 10.1006/expr.1998.4346

M3 - Article

C2 - 9806873

AN - SCOPUS:0032212406

SN - 0014-4894

VL - 90

SP - 277

EP - 285

JO - Experimental Parasitology

JF - Experimental Parasitology

IS - 3

ER -

Neospora caninum: Tachyzoites express a potent type-I nucleoside triphosphate hydrolase, but lack nucleoside diphosphate hydrolase activity

Abstract

Bibliographical note

Funding

Keywords

ASJC Scopus subject areas

UN SDGs

Access to Document

Other files and links

Fingerprint

Cite this