PMID- 31775627
OWN - NLM
STAT- MEDLINE
DCOM- 20200408
LR  - 20200408
IS  - 2661-8850 (Electronic)
IS  - 2661-8850 (Linking)
VI  - 20
IP  - 1
DP  - 2019 Nov 27
TI  - Detection of 8-oxoguanine and apurinic/apyrimidinic sites using a 
      fluorophore-labeled probe with cell-penetrating ability.
PG  - 54
LID - 10.1186/s12860-019-0236-x [doi]
LID - 54
AB  - BACKGROUND: Reactive oxygen species (ROS) produce different lesions in DNA by 
      ROS-induced DNA damage. Detection and quantification of 8-oxo-7,8-dihydroguanine 
      (8-oxoG) within cells are important for study. Human ribosomal protein S3 (hRpS3) 
      has a high binding affinity to 8-oxoG. In this study, we developed an imaging 
      probe to detect 8-oxoG using a specific peptide from hRpS3. Transactivator (TAT) 
      proteins are known to have cell-penetrating properties. Therefore, we developed a 
      TAT-S3 probe by attaching a TAT peptide to our imaging probe. RESULTS: A DNA 
      binding assay was conducted to confirm that our probe bound to 8-oxoG and 
      apurinic/apyrimidinic (AP) sites. We confirmed that the TAT-S3 probe localized in 
      the mitochondria, without permeabilization, and fluoresced in H(2)O(2)-treated 
      HeLa cells and zebrafish embryos. Treatment with Mitoquinone (MitoQ), a 
      mitochondria-targeted antioxidant, reduced TAT-S3 probe fluorescence. 
      Additionally, treatment with O8, an inhibitor of OGG1, increased probe 
      fluorescence. A competition assay was conducted with an aldehyde reaction probe 
      (ARP) and methoxyamine (MX) to confirm binding of TAT-S3 to the AP sites. The 
      TAT-S3 probe showed competitive binding to AP sites with ARP and MX. CONCLUSIONS: 
      These results revealed that the TAT-S3 probe successfully detected the presence 
      of 8-oxoG and AP sites in damaged cells. The TAT-S3 probe may have applications 
      for the detection of diseases caused by reactive oxygen species.
FAU - Kang, Dong Min
AU  - Kang DM
AD  - Department of Advanced Technology Fusion, Konkuk University, 120 Neungdong-ro, 
      Gwangjin-gu, Seoul, 05029, Republic of Korea.
FAU - Shin, Jong-Il
AU  - Shin JI
AD  - Department of Biological Sciences, Konkuk University, Seoul, 05029, South Korea.
FAU - Kim, Ji Beom
AU  - Kim JB
AD  - Department of Biological Sciences, Konkuk University, Seoul, 05029, South Korea.
FAU - Lee, Kyungho
AU  - Lee K
AD  - Department of Biological Sciences, Konkuk University, Seoul, 05029, South Korea.
FAU - Chung, Ji Hyung
AU  - Chung JH
AD  - Department of Applied Bioscience, College of Life Science, CHA University, 
      Pocheon, 11160, South Korea.
FAU - Yang, Hye-Won
AU  - Yang HW
AD  - Department of Marine Life Science, Jeju National University, Jeju, 63243, South 
      Korea.
FAU - Kim, Kil-Nam
AU  - Kim KN
AD  - Chuncheon Center, Korea Basic Science Institute (KBSI), Chuncheon, 24341, South 
      Korea.
FAU - Han, Ye Sun
AU  - Han YS
AUID- ORCID: 0000-0001-7214-1513
AD  - Department of Advanced Technology Fusion, Konkuk University, 120 Neungdong-ro, 
      Gwangjin-gu, Seoul, 05029, Republic of Korea. yshan@konkuk.ac.kr.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20191127
PL  - England
TA  - BMC Mol Cell Biol
JT  - BMC molecular and cell biology
JID - 101741148
RN  - 0 (DNA, Mitochondrial)
RN  - 0 (Fluorescent Dyes)
RN  - 0 (RPS3 protein, human)
RN  - 0 (Ribosomal Proteins)
RN  - 0 (Trans-Activators)
RN  - 5614-64-2 (8-hydroxyguanine)
RN  - 5Z93L87A1R (Guanine)
RN  - 9007-49-2 (DNA)
RN  - EC 4.2.99.18 (DNA-(Apurinic or Apyrimidinic Site) Lyase)
SB  - IM
MH  - Animals
MH  - Binding Sites
MH  - DNA/*analysis/chemistry
MH  - DNA Damage
MH  - DNA, Mitochondrial
MH  - DNA-(Apurinic or Apyrimidinic Site) Lyase/analysis/chemistry
MH  - Flow Cytometry
MH  - *Fluorescent Dyes/chemical synthesis
MH  - Guanine/*analogs & derivatives/analysis/metabolism
MH  - HeLa Cells
MH  - Humans
MH  - Microscopy, Confocal
MH  - Mitochondria/pathology
MH  - Protein Binding
MH  - Ribosomal Proteins/chemistry/metabolism
MH  - Trans-Activators/chemistry
MH  - Zebrafish
PMC - PMC6881995
OTO - NOTNLM
OT  - 8-oxo-7,8-dihydroguanine (8-oxoG)
OT  - Apurinic/apyrimidinic (AP) site
OT  - Human ribosomal protein S3 (hRpS3)
OT  - Transactivator (TAT) proteins
COIS- The authors declare that they have no competing interests.
EDAT- 2019/11/30 06:00
MHDA- 2020/04/09 06:00
PMCR- 2019/11/27
CRDT- 2019/11/29 06:00
PHST- 2019/06/13 00:00 [received]
PHST- 2019/11/14 00:00 [accepted]
PHST- 2019/11/29 06:00 [entrez]
PHST- 2019/11/30 06:00 [pubmed]
PHST- 2020/04/09 06:00 [medline]
PHST- 2019/11/27 00:00 [pmc-release]
AID - 10.1186/s12860-019-0236-x [pii]
AID - 236 [pii]
AID - 10.1186/s12860-019-0236-x [doi]
PST - epublish
SO  - BMC Mol Cell Biol. 2019 Nov 27;20(1):54. doi: 10.1186/s12860-019-0236-x.