Regulation of 92 kDa type IV collagenase expression by the jun aminoterminal kinase- and the extracellular signal-regulated kinase-dependent signaling cascades View Full Text


Ontology type: schema:ScholarlyArticle      Open Access: True


Article Info

DATE

1997-03

AUTHORS

R Gum, H Wang, E Lengyel, J Juarez, D Boyd

ABSTRACT

The 92 kDa type IV collagenase (MMP-9), which degrades type IV collagen, has been implicated in tissue remodeling. The purpose of the current study was to determine the role of Jun amino-terminal kinase (JNK)- and extracellular signal-regulated kinase- (ERK)-dependent signaling cascades in the regulation of MMP-9 expression. Towards this end, we first determined the transcriptional requirements for MMP-9 promoter activity in a cell line (UM-SCC-1) which is an avid secretor of this collagenase. Transfection of these cells with a CAT reporter driven by progressive 5′ deleted fragments of the MMP-9 promoter indicated the requirement of a region spanning −144 to −73 for optimal promoter activity. DNase I footprinting revealed a protected region of the promoter spanning nucleotides −91 to −68 and containing a consensus AP-1 motif at −79. Mutation of this AP-1 motif practically abolished the activity of the MMP-9 promoter-driven CAT reporter. Mobility shift assays indicated c-Fos and Jun-D bound to this motif and transfection of the cells with a mutated c-Jun, which quenches the function of endogenous Jun and Fos proteins, decreased MMP-9 promoter activity by 80%. UM-SCC-1 cells contained a constitutively activated JNK and the expression of a kinase-deficient JNK1 reduced the activity of a CAT reporter driven either by the MMP-9 promoter or by three tandem AP-1 repeats upstream of a thymidine kinase minimal promoter. Conditioned medium collected from UM-SCC-1 cells transfected with the dominant negative JNK1 expression vector diminished 92 kDa gelatinolysis. Similarly, interfering with MEKK, which lies upstream of JNK1, using a dominant negative expression vector reduced MMP-9 promoter activity over the same concentration range which repressed the AP-1-thymidine kinase CAT reporter construct. UM-SCC-1 cells also contained a constitutively activated ERK1. MMP-9 expression, as determined by CAT assays and by zymography, was reduced by the co-expression of a kinase-deficient ERK1. Interfering with MEK1, which is an upstream activator of ERK1, either with PD 098059, which prevents the activation of MEK1, or with a dominant negative expression construct, reduced 92 kDa gelatinolysis and MMP-9 promoter activity respectively. c-Raf-1 is an upstream activator of MEK1 and a kinase-deficient c-Raf-1 expression construct decreased the activity of a promoter driven by either the MMP-9 promoter or three tandem AP-1 repeats. Conversely, treatment of UM-SCC-1 cells with PMA, which activates c-Raf-1, increased 92 kDa gelatinolysis. These data suggest that MMP-9 expression in UM-SCC-1 cells, is regulated by JNK- and ERK-dependent signaling pathways. More... »

PAGES

1481-1493

Journal

TITLE

Oncogene

ISSUE

12

VOLUME

14

Identifiers

URI

http://scigraph.springernature.com/pub.10.1038/sj.onc.1200973

DOI

http://dx.doi.org/10.1038/sj.onc.1200973

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1027441441

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/9136992


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75 avid secretor
76 cascade
77 cell lines
78 cells
79 collagen
80 collagenase
81 collagenase expression
82 concentration range
83 consensus AP-1 motif
84 constructs
85 current study
86 data
87 dominant negative JNK1 expression vector
88 dominant negative expression constructs
89 dominant negative expression vectors
90 end
91 endogenous Jun
92 expression
93 expression constructs
94 expression vector
95 extracellular signal-regulated kinase
96 extracellular signal-regulated kinase-dependent signaling cascades
97 fragments
98 function
99 gelatinolysis
100 kDa type IV collagenase expression
101 kinase
102 kinase CAT reporter construct
103 kinase minimal promoter
104 kinase-deficient ERK1
105 kinase-deficient JNK1
106 kinase-dependent signaling cascades
107 lines
108 medium
109 minimal promoter
110 mobility shift
111 motif
112 mutations
113 negative JNK1 expression vector
114 negative expression construct
115 negative expression vector
116 nucleotides
117 optimal promoter activity
118 pathway
119 promoter
120 promoter activity
121 promoter spanning nucleotides
122 promoter-driven CAT reporter
123 protein
124 purpose
125 range
126 region
127 regulation
128 reporter
129 reporter constructs
130 requirements
131 role
132 same concentration range
133 secretors
134 shift
135 signal-regulated kinase
136 signal-regulated kinase-dependent signaling cascades
137 signaling cascades
138 signaling pathways
139 spanning nucleotides
140 study
141 tandem AP-1
142 thymidine kinase minimal promoter
143 tissue
144 transcriptional requirements
145 transfection
146 treatment
147 type IV collagen
148 type IV collagenase
149 type IV collagenase expression
150 upstream activator
151 vector
152 zymography
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