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ADP-Ribosylation Factor 6 Expression and Activation Are Reduced in Myometrium in Complicated Pregnancies

Venkat Kanamarlapudi Orcid Logo, Sian-eleri Owens Orcid Logo, Jon Lartey, Andres Lopez Bernal

PLoS ONE, Volume: 7, Issue: 5, Start page: e37954

Swansea University Authors: Venkat Kanamarlapudi Orcid Logo, Sian-eleri Owens Orcid Logo

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DOI (Published version): 10.1371/journal.pone.0037954

Abstract

BACKGROUND:ARF6 (ADP-ribosylation factor 6) small GTP binding protein plays critical roles in actin cytoskeleton rearrangements and membrane trafficking, including internalisation of G protein coupled receptors (GPCR). ARF6 operates by cycling between GDP-bound (inactive) and GTP-bound (active) form...

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Published in: PLoS ONE
ISSN: 1932-6203
Published: 2012
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URI: https://cronfa.swan.ac.uk/Record/cronfa11278
first_indexed 2013-07-23T12:05:18Z
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spelling 2022-02-28T12:48:03.3659726 v2 11278 2012-06-12 ADP-Ribosylation Factor 6 Expression and Activation Are Reduced in Myometrium in Complicated Pregnancies 63741801137148abfa4c00cd547dcdfa 0000-0002-8739-1483 Venkat Kanamarlapudi Venkat Kanamarlapudi true false 721deb4604d122019244cfdf08820cbe 0000-0003-1806-5235 Sian-eleri Owens Sian-eleri Owens true false 2012-06-12 MEDS BACKGROUND:ARF6 (ADP-ribosylation factor 6) small GTP binding protein plays critical roles in actin cytoskeleton rearrangements and membrane trafficking, including internalisation of G protein coupled receptors (GPCR). ARF6 operates by cycling between GDP-bound (inactive) and GTP-bound (active) forms and is a potential regulator of GPCR-mediated uterine activity during pregnancy and labour. ARF6 contains very low intrinsic GTP binding activity and depends on GEFs (guanine nucleotide exchange factors) such as CYTH3 (cytohesin 3) to bind GTP. ARF6 and CYTH3 were originally cloned from human placenta, but there is no information on their expression in other reproductive tissues.METHODS:The expression of ARF6, ARF1, and CYTH1-4 was investigated by measuring mRNA (using RT-PCR) and protein levels (using immunoblotting) in samples of myometrium obtained from non-pregnant women, and women with normal pregnancies, before or after the spontaneous onset of labour. We also analysed myometrial samples from women with spontaneous preterm labour and from women with complicated pregnancies requiring emergency preterm delivery. The GST)-effector pull down assay was used to study the presence of active ARF6 and ARF1 in all myometrial extracts.RESULTS:ARF6, ARF1 and CYTH3 but not CYTH1, CYTH2 and CYTH4 were expressed in all samples and the levels did not change with pregnancy or labour. However, ARF6 and CYTH3 but not ARF1 levels were significantly reduced in complicated pregnancies. The alterations in the expression of ARF6 and its GEF in human myometrium indicate a potential involvement of this signalling system in modulating the response of myometrial smooth muscle in complicated pregnancies. The levels of ARF6-GTP or ARF1-GTP did not change with pregnancy or labour but ARF6-GTP levels were significantly decreased in women with severe complications of pregnancy.CONCLUSIONS:We have demonstrated a functional ARF6 system in human myometrium and a correlation between ARF6 level and activity in uterine and abnormal pregnancy. Journal Article PLoS ONE 7 5 e37954 1932-6203 30 5 2012 2012-05-30 10.1371/journal.pone.0037954 http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.0037954 COLLEGE NANME Medical School COLLEGE CODE MEDS Swansea University 2022-02-28T12:48:03.3659726 2012-06-12T19:29:09.7108053 Faculty of Medicine, Health and Life Sciences Swansea University Medical School - Medicine Venkat Kanamarlapudi 0000-0002-8739-1483 1 Sian-eleri Owens 0000-0003-1806-5235 2 Jon Lartey 3 Andres Lopez Bernal 4 0011278-16072019135542.pdf ARF6expmyometriumpregnanPLoSONE12v2.pdf 2019-07-16T13:55:42.1300000 Output 807632 application/pdf Not Applicable (or Unknown) true 2019-06-16T00:00:00.0000000 false eng
title ADP-Ribosylation Factor 6 Expression and Activation Are Reduced in Myometrium in Complicated Pregnancies
spellingShingle ADP-Ribosylation Factor 6 Expression and Activation Are Reduced in Myometrium in Complicated Pregnancies
Venkat Kanamarlapudi
Sian-eleri Owens
title_short ADP-Ribosylation Factor 6 Expression and Activation Are Reduced in Myometrium in Complicated Pregnancies
title_full ADP-Ribosylation Factor 6 Expression and Activation Are Reduced in Myometrium in Complicated Pregnancies
title_fullStr ADP-Ribosylation Factor 6 Expression and Activation Are Reduced in Myometrium in Complicated Pregnancies
title_full_unstemmed ADP-Ribosylation Factor 6 Expression and Activation Are Reduced in Myometrium in Complicated Pregnancies
title_sort ADP-Ribosylation Factor 6 Expression and Activation Are Reduced in Myometrium in Complicated Pregnancies
author_id_str_mv 63741801137148abfa4c00cd547dcdfa
721deb4604d122019244cfdf08820cbe
author_id_fullname_str_mv 63741801137148abfa4c00cd547dcdfa_***_Venkat Kanamarlapudi
721deb4604d122019244cfdf08820cbe_***_Sian-eleri Owens
author Venkat Kanamarlapudi
Sian-eleri Owens
author2 Venkat Kanamarlapudi
Sian-eleri Owens
Jon Lartey
Andres Lopez Bernal
format Journal article
container_title PLoS ONE
container_volume 7
container_issue 5
container_start_page e37954
publishDate 2012
institution Swansea University
issn 1932-6203
doi_str_mv 10.1371/journal.pone.0037954
college_str Faculty of Medicine, Health and Life Sciences
hierarchytype
hierarchy_top_id facultyofmedicinehealthandlifesciences
hierarchy_top_title Faculty of Medicine, Health and Life Sciences
hierarchy_parent_id facultyofmedicinehealthandlifesciences
hierarchy_parent_title Faculty of Medicine, Health and Life Sciences
department_str Swansea University Medical School - Medicine{{{_:::_}}}Faculty of Medicine, Health and Life Sciences{{{_:::_}}}Swansea University Medical School - Medicine
url http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.0037954
document_store_str 1
active_str 0
description BACKGROUND:ARF6 (ADP-ribosylation factor 6) small GTP binding protein plays critical roles in actin cytoskeleton rearrangements and membrane trafficking, including internalisation of G protein coupled receptors (GPCR). ARF6 operates by cycling between GDP-bound (inactive) and GTP-bound (active) forms and is a potential regulator of GPCR-mediated uterine activity during pregnancy and labour. ARF6 contains very low intrinsic GTP binding activity and depends on GEFs (guanine nucleotide exchange factors) such as CYTH3 (cytohesin 3) to bind GTP. ARF6 and CYTH3 were originally cloned from human placenta, but there is no information on their expression in other reproductive tissues.METHODS:The expression of ARF6, ARF1, and CYTH1-4 was investigated by measuring mRNA (using RT-PCR) and protein levels (using immunoblotting) in samples of myometrium obtained from non-pregnant women, and women with normal pregnancies, before or after the spontaneous onset of labour. We also analysed myometrial samples from women with spontaneous preterm labour and from women with complicated pregnancies requiring emergency preterm delivery. The GST)-effector pull down assay was used to study the presence of active ARF6 and ARF1 in all myometrial extracts.RESULTS:ARF6, ARF1 and CYTH3 but not CYTH1, CYTH2 and CYTH4 were expressed in all samples and the levels did not change with pregnancy or labour. However, ARF6 and CYTH3 but not ARF1 levels were significantly reduced in complicated pregnancies. The alterations in the expression of ARF6 and its GEF in human myometrium indicate a potential involvement of this signalling system in modulating the response of myometrial smooth muscle in complicated pregnancies. The levels of ARF6-GTP or ARF1-GTP did not change with pregnancy or labour but ARF6-GTP levels were significantly decreased in women with severe complications of pregnancy.CONCLUSIONS:We have demonstrated a functional ARF6 system in human myometrium and a correlation between ARF6 level and activity in uterine and abnormal pregnancy.
published_date 2012-05-30T18:20:55Z
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score 11.04748

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