PMID- 23216940
OWN - NLM
STAT- MEDLINE
DCOM- 20130701
LR  - 20220408
IS  - 1477-7827 (Electronic)
IS  - 1477-7827 (Linking)
VI  - 10
DP  - 2012 Dec 5
TI  - Myeloid cell leukemia-1 (Mc1-1) is a candidate target gene of hypoxia-inducible 
      factor-1 (HIF-1) in the testis.
PG  - 104
LID - 10.1186/1477-7827-10-104 [doi]
AB  - BACKGROUND: Spermatic cord torsion can lead to testis ischemia (I) and subsequent 
      ischemia-reperfusion (I/R) causing germ cell-specific apoptosis. Previously, we 
      demonstrated that the hypoxia-inducible factor-1 (HIF-1) transcription factor, a 
      key regulator of physiological responses to hypoxia, is abundant in Leydig cells 
      in normoxic and ischemic testes. We hypothesize that testicular HIF-1 activates 
      the expression of antiapoptotic target genes to protect Leydig cells from 
      apoptosis. In silico analysis of testis genes containing a consensus hypoxia 
      response element (HRE, 5'-RCGTG-3') identified myeloid cell leukemia-1 (Mcl-1) as 
      a potential HIF-1 target gene. The purpose of this study was to determine whether 
      HIF-1 shows DNA-binding activity in normoxic and ischemic testes and whether 
      Mcl-1 is a target gene of testicular HIF-1. METHODS: The testicular HIF-1 
      DNA-binding capacity was analyzed in vitro using a quantitative enzyme-linked 
      immunosorbent assay (ELISA) and electrophoretic mobility shift assays (EMSA). 
      MCL-1 protein expression was evaluated by immunoblot analysis and 
      immunohistochemistry. The binding of testicular HIF-1 to the Mcl-1 gene was 
      examined via chromatin immunoprecipitation (ChIP) analysis. RESULTS: The ELISA 
      and EMSA assays demonstrated that testicular HIF-1 from normoxic and ischemic 
      testes binds DNA equally strongly, suggesting physiological roles for HIF-1 in 
      the normoxic testis, unlike most tissues in which HIF-1 is degraded under 
      normoxic conditions and is only activated by hypoxia. MCL-1 protein was 
      determined to be abundant in both normoxic and ischemic testes and expressed in 
      Leydig cells. In a pattern identical to that of HIF-1 expression, the 
      steady-state levels of MCL-1 were not significantly affected by I or I/R and 
      MCL-1 co-localized with HIF-1alpha in Leydig cells. Chromatin immunoprecipitation 
      (ChIP) analysis using a HIF-1 antibody revealed sequences enriched for the Mcl-1 
      promoter. CONCLUSIONS: The results demonstrated that, unlike what is observed in 
      most tissues, HIF-1 displays DNA-binding activity in both normoxic and ischemic 
      testes, and Mcl-1 may be a key target gene of testicular HIF-1 with potential 
      roles in the antiapoptotic protection of Leydig cells.
FAU - Palladino, Michael A
AU  - Palladino MA
AD  - Department of Biology, Monmouth University, 400 Cedar Avenue, West Long Branch, 
      NJ 07764, USA. mpalladi@monmouth.edu
FAU - Shah, Anoop
AU  - Shah A
FAU - Tyson, Rebecca
AU  - Tyson R
FAU - Horvath, Jaclyn
AU  - Horvath J
FAU - Dugan, Christine
AU  - Dugan C
FAU - Karpodinis, Marie
AU  - Karpodinis M
LA  - eng
GR  - R15-HD046451/HD/NICHD NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
DEP - 20121205
PL  - England
TA  - Reprod Biol Endocrinol
JT  - Reproductive biology and endocrinology : RB&E
JID - 101153627
RN  - 0 (Hypoxia-Inducible Factor 1)
RN  - 0 (Hypoxia-Inducible Factor 1, alpha Subunit)
RN  - 0 (Myeloid Cell Leukemia Sequence 1 Protein)
RN  - 0 (Proto-Oncogene Proteins c-bcl-2)
RN  - 9007-49-2 (DNA)
SB  - IM
MH  - Animals
MH  - Apoptosis
MH  - Cell Hypoxia
MH  - Chromatin Immunoprecipitation
MH  - DNA/metabolism
MH  - Hypoxia-Inducible Factor 1/*analysis/*metabolism
MH  - Hypoxia-Inducible Factor 1, alpha Subunit/analysis
MH  - Immunoblotting
MH  - Immunohistochemistry
MH  - Ischemia/metabolism
MH  - Leydig Cells/chemistry
MH  - Male
MH  - Myeloid Cell Leukemia Sequence 1 Protein
MH  - Proto-Oncogene Proteins c-bcl-2/analysis/*genetics
MH  - Rats
MH  - Rats, Sprague-Dawley
MH  - Testis/blood supply/*chemistry
PMC - PMC3556106
EDAT- 2012/12/12 06:00
MHDA- 2013/07/03 06:00
PMCR- 2012/12/05
CRDT- 2012/12/11 06:00
PHST- 2012/08/08 00:00 [received]
PHST- 2012/11/27 00:00 [accepted]
PHST- 2012/12/11 06:00 [entrez]
PHST- 2012/12/12 06:00 [pubmed]
PHST- 2013/07/03 06:00 [medline]
PHST- 2012/12/05 00:00 [pmc-release]
AID - 1477-7827-10-104 [pii]
AID - 10.1186/1477-7827-10-104 [doi]
PST - epublish
SO  - Reprod Biol Endocrinol. 2012 Dec 5;10:104. doi: 10.1186/1477-7827-10-104.