Novel GC-rich DNA-binding compound produced by a genetically engineered mutant of the mithramycin producer Streptomyces argillaceus exhibits improved transcriptional repressor activity: implications for cancer therapy.
The aureolic acid antibiotic mithramycin (MTM) binds selectively to GC-rich DNA sequences and blocks preferentially binding of proteins, like Sp1 transcription factors, to GC-rich elements in gene promoters. Genetic approaches can be applied to alter the MTM biosynthetic pathway in the producing microorganism and obtain new products with improved pharmacological properties. Here, we report on a new analog, MTM SDK, obtained by targeted gene inactivation of the ketoreductase MtmW catalyzing the last step in MTM biosynthesis. SDK exhibited greater activity as transcriptional inhibitor compared to MTM. SDK was a potent inhibitor of Sp1-dependent reporter activity and interfered minimally with reporters of other transcription factors, indicating that it retained a high degree of selectivity toward GC-rich DNA-binding transcription factors. RT-PCR and microarray analysis showed that SDK repressed transcription of multiple genes implicated in critical aspects of cancer development and progression, including cell cycle, apoptosis, migration, invasion and angiogenesis, consistent with the pleiotropic role of Sp1 family transcription factors. SDK inhibited proliferation and was a potent inducer of apoptosis in ovarian cancer cells while it had minimal effects on viability of normal cells. The new MTM derivative SDK could be an effective agent for treatment of cancer and other diseases with abnormal expression or activity of GC-rich DNA-binding transcription factors.
Laboratory of Experimental Oncology, Oncology Institute of Southern Switzerland Bellinzona, Switzerland.
Veronica
V
Albertini
Aklank
A
Jain
Sara
S
Vignati
Sara
S
Napoli
Andrea
A
Rinaldi
Ivo
I
Kwee
Mohammad
M
Nur-e-Alam
Julia
J
Bergant
Francesco
F
Bertoni
Giuseppina M
GM
Carbone
Jürgen
J
Rohr
Carlo V
CV
Catapano
D000903:Antibiotics, Antineoplastic / Q000737:chemistry / Q000378:metabolism; D000970:Antineoplastic Agents / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D017209:Apoptosis; D002453:Cell Cycle / Q000187:drug effects; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D004247:DNA / Q000737:chemistry / Q000378:metabolism; D005260:Female; D020862:GC Rich Sequence; D020869:Gene Expression Profiling; D020868:Gene Silencing; D005818:Genetic Engineering; D006801:Humans; D009154:Mutation; D009369:Neoplasms / Q000188:drug therapy; D010051:Ovarian Neoplasms / Q000473:pathology; D010088:Oxidoreductases / Q000235:genetics; D008926:Plicamycin / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D013302:Streptomyces / Q000235:genetics* / Q000378:metabolism; D014158:Transcription, Genetic / Q000187:drug effects*
D000903:Antibiotics, Antineoplastic; D000970:Antineoplastic Agents; C509194:SDK compound; D004247:DNA; D010088:Oxidoreductases; D008926:Plicamycin