Identification of Apurinic/apyrimidinic endonuclease 1 (APE1) as the endoribonuclease that cleaves c-myc mRNA

Tavish Barnes; Wan Cheol Kim; Anil K. Mantha; Sang Eun Kim; Tadahide Izumi; Sankar Mitra; Chow H. Lee

doi:10.1093/nar/gkp275

Identification of Apurinic/apyrimidinic endonuclease 1 (APE1) as the endoribonuclease that cleaves c-myc mRNA

Tavish Barnes, Wan Cheol Kim, Anil K. Mantha, Sang Eun Kim, Tadahide Izumi, Sankar Mitra, Chow H. Lee

Toxicology and Cancer Biology

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

125 Scopus citations

Abstract

Endonucleolytic cleavage of the coding region determinant (CRD) of c-myc mRNA appears to play a critical role in regulating c-myc mRNA turnover. Using ³²P-labeled c-myc CRD RNA as substrate, we have purified and identified two endoribonucleases from rat liver polysomes that are capable of cleaving the transcript in vitro. A 17-kDa enzyme was identified as RNase1. Apurinic/apyrimidinic (AP) DNA endonuclease 1 (APE1) was identified as the 35-kDa endoribonuclease that preferentially cleaves in between UA and CA dinucleotides of c-myc CRD RNA. APE1 was further confirmed to be the 35-kDa endoribonuclease because: (i) the endoribonuclease activity of the purified 35-kDa native enzyme was specifically immuno-depleted with APE1 monoclonal antibody, and (ii) recombinant human APE1 generated identical RNA cleavage patterns as the native liver enzyme. Studies using E96A and H309N mutants of APE1 suggest that the endoribonuclease activity for c-myc CRD RNA shares the same active center with the AP-DNA endonuclease activity. Transient knockdown of APE1 in HeLa cells led to increased steady-state level of c-myc mRNA and its half-life. We conclude that the ability to cleave RNA dinucleotides is a previously unidentified function of APE1 and it can regulate c-myc mRNA level possibly via its endoribonuclease activity.

Original language	English
Pages (from-to)	3946-3958
Number of pages	13
Journal	Nucleic Acids Research
Volume	37
Issue number	12
DOIs	https://doi.org/10.1093/nar/gkp275
State	Published - 2009

Bibliographical note

Funding Information:
Natural Sciences & Engineering Research Council (227158); the Canadian Cancer Society [to C.H.L, who is a Research Scientist of the National Cancer Institute of Canada (13546)]; T.B was a recipient of UNBC Graduate Entrance and NSERC Canada Graduate Scholarships; W-C.K was a recipient of Pacific Century, Michael Smith Foundation of Health Research Junior Graduate, and NSERC Canada Graduate Scholarships; S-E.K was a recipient of NSERC Undergraduate Summer Research Award; the United States Public Health Service grants (R01 ES08457, R01 CA53791, and P30 ES06676 to S.M. and A.K.M.); and National Institutes of Health/National Cancer Institute grant (CA98664 to T.I.). Funding for open access charge: Natural Sciences & Engineering Research Council Discovery Grant 227158.

ASJC Scopus subject areas

Genetics

Access to Document

10.1093/nar/gkp275

Cite this

@article{463bd558418d4aa0ac98315bc2827ff3,

title = "Identification of Apurinic/apyrimidinic endonuclease 1 (APE1) as the endoribonuclease that cleaves c-myc mRNA",

abstract = "Endonucleolytic cleavage of the coding region determinant (CRD) of c-myc mRNA appears to play a critical role in regulating c-myc mRNA turnover. Using 32P-labeled c-myc CRD RNA as substrate, we have purified and identified two endoribonucleases from rat liver polysomes that are capable of cleaving the transcript in vitro. A 17-kDa enzyme was identified as RNase1. Apurinic/apyrimidinic (AP) DNA endonuclease 1 (APE1) was identified as the 35-kDa endoribonuclease that preferentially cleaves in between UA and CA dinucleotides of c-myc CRD RNA. APE1 was further confirmed to be the 35-kDa endoribonuclease because: (i) the endoribonuclease activity of the purified 35-kDa native enzyme was specifically immuno-depleted with APE1 monoclonal antibody, and (ii) recombinant human APE1 generated identical RNA cleavage patterns as the native liver enzyme. Studies using E96A and H309N mutants of APE1 suggest that the endoribonuclease activity for c-myc CRD RNA shares the same active center with the AP-DNA endonuclease activity. Transient knockdown of APE1 in HeLa cells led to increased steady-state level of c-myc mRNA and its half-life. We conclude that the ability to cleave RNA dinucleotides is a previously unidentified function of APE1 and it can regulate c-myc mRNA level possibly via its endoribonuclease activity.",

author = "Tavish Barnes and Kim, {Wan Cheol} and Mantha, {Anil K.} and Kim, {Sang Eun} and Tadahide Izumi and Sankar Mitra and Lee, {Chow H.}",

note = "Funding Information: Natural Sciences & Engineering Research Council (227158); the Canadian Cancer Society [to C.H.L, who is a Research Scientist of the National Cancer Institute of Canada (13546)]; T.B was a recipient of UNBC Graduate Entrance and NSERC Canada Graduate Scholarships; W-C.K was a recipient of Pacific Century, Michael Smith Foundation of Health Research Junior Graduate, and NSERC Canada Graduate Scholarships; S-E.K was a recipient of NSERC Undergraduate Summer Research Award; the United States Public Health Service grants (R01 ES08457, R01 CA53791, and P30 ES06676 to S.M. and A.K.M.); and National Institutes of Health/National Cancer Institute grant (CA98664 to T.I.). Funding for open access charge: Natural Sciences & Engineering Research Council Discovery Grant 227158.",

year = "2009",

doi = "10.1093/nar/gkp275",

language = "English",

volume = "37",

pages = "3946--3958",

number = "12",

}

TY - JOUR

T1 - Identification of Apurinic/apyrimidinic endonuclease 1 (APE1) as the endoribonuclease that cleaves c-myc mRNA

AU - Barnes, Tavish

AU - Kim, Wan Cheol

AU - Mantha, Anil K.

AU - Kim, Sang Eun

AU - Izumi, Tadahide

AU - Mitra, Sankar

AU - Lee, Chow H.

N1 - Funding Information: Natural Sciences & Engineering Research Council (227158); the Canadian Cancer Society [to C.H.L, who is a Research Scientist of the National Cancer Institute of Canada (13546)]; T.B was a recipient of UNBC Graduate Entrance and NSERC Canada Graduate Scholarships; W-C.K was a recipient of Pacific Century, Michael Smith Foundation of Health Research Junior Graduate, and NSERC Canada Graduate Scholarships; S-E.K was a recipient of NSERC Undergraduate Summer Research Award; the United States Public Health Service grants (R01 ES08457, R01 CA53791, and P30 ES06676 to S.M. and A.K.M.); and National Institutes of Health/National Cancer Institute grant (CA98664 to T.I.). Funding for open access charge: Natural Sciences & Engineering Research Council Discovery Grant 227158.

PY - 2009

Y1 - 2009

N2 - Endonucleolytic cleavage of the coding region determinant (CRD) of c-myc mRNA appears to play a critical role in regulating c-myc mRNA turnover. Using 32P-labeled c-myc CRD RNA as substrate, we have purified and identified two endoribonucleases from rat liver polysomes that are capable of cleaving the transcript in vitro. A 17-kDa enzyme was identified as RNase1. Apurinic/apyrimidinic (AP) DNA endonuclease 1 (APE1) was identified as the 35-kDa endoribonuclease that preferentially cleaves in between UA and CA dinucleotides of c-myc CRD RNA. APE1 was further confirmed to be the 35-kDa endoribonuclease because: (i) the endoribonuclease activity of the purified 35-kDa native enzyme was specifically immuno-depleted with APE1 monoclonal antibody, and (ii) recombinant human APE1 generated identical RNA cleavage patterns as the native liver enzyme. Studies using E96A and H309N mutants of APE1 suggest that the endoribonuclease activity for c-myc CRD RNA shares the same active center with the AP-DNA endonuclease activity. Transient knockdown of APE1 in HeLa cells led to increased steady-state level of c-myc mRNA and its half-life. We conclude that the ability to cleave RNA dinucleotides is a previously unidentified function of APE1 and it can regulate c-myc mRNA level possibly via its endoribonuclease activity.

AB - Endonucleolytic cleavage of the coding region determinant (CRD) of c-myc mRNA appears to play a critical role in regulating c-myc mRNA turnover. Using 32P-labeled c-myc CRD RNA as substrate, we have purified and identified two endoribonucleases from rat liver polysomes that are capable of cleaving the transcript in vitro. A 17-kDa enzyme was identified as RNase1. Apurinic/apyrimidinic (AP) DNA endonuclease 1 (APE1) was identified as the 35-kDa endoribonuclease that preferentially cleaves in between UA and CA dinucleotides of c-myc CRD RNA. APE1 was further confirmed to be the 35-kDa endoribonuclease because: (i) the endoribonuclease activity of the purified 35-kDa native enzyme was specifically immuno-depleted with APE1 monoclonal antibody, and (ii) recombinant human APE1 generated identical RNA cleavage patterns as the native liver enzyme. Studies using E96A and H309N mutants of APE1 suggest that the endoribonuclease activity for c-myc CRD RNA shares the same active center with the AP-DNA endonuclease activity. Transient knockdown of APE1 in HeLa cells led to increased steady-state level of c-myc mRNA and its half-life. We conclude that the ability to cleave RNA dinucleotides is a previously unidentified function of APE1 and it can regulate c-myc mRNA level possibly via its endoribonuclease activity.

UR - http://www.scopus.com/inward/record.url?scp=67651149733&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=67651149733&partnerID=8YFLogxK

U2 - 10.1093/nar/gkp275

DO - 10.1093/nar/gkp275

M3 - Article

C2 - 19401441

AN - SCOPUS:67651149733

SN - 0305-1048

VL - 37

SP - 3946

EP - 3958

JO - Nucleic Acids Research

JF - Nucleic Acids Research

IS - 12

ER -

Identification of Apurinic/apyrimidinic endonuclease 1 (APE1) as the endoribonuclease that cleaves c-myc mRNA

Abstract

Bibliographical note

ASJC Scopus subject areas

Access to Document

Other files and links

Fingerprint

Cite this