Tetratricopeptide repeat domain 9A is an interacting protein for tropomyosin Tm5NM-1.
BACKGROUND
Tetratricopeptide repeat domain 9A (TTC9A) protein is a recently identified protein which contains three tetratricopeptide repeats (TPRs) on its C-terminus. In our previous studies, we have shown that TTC9A was a hormonally-regulated gene in breast cancer cells. In this study, we found that TTC9A was over-expressed in breast cancer tissues compared with the adjacent controls (P < 0.00001), suggesting it might be involved in the breast cancer development process. The aim of the current study was to further elucidate the function of TTC9A.


METHODS
Breast samples from 25 patients including the malignant breast tissues and the adjacent normal tissues were processed for Southern blot analysis. Yeast-two-hybrid assay, GST pull-down assay and co-immunoprecipitation were used to identify and verify the interaction between TTC9A and other proteins.


RESULTS
Tropomyosin Tm5NM-1 was identified as one of the TTC9A partner proteins. The interaction between TTC9A and Tm5NM-1 was further confirmed by GST pull-down assay and co-immunoprecipitation in mammalian cells. TTC9A domains required for the interaction were also characterized in this study. The results suggested that the first TPR domain and the linker fragment between the first two TPR domains of TTC9A were important for the interaction with Tm5NM-1 and the second and the third TPR might play an inhibitory role.


CONCLUSIONS
Since the primary function of tropomyosin is to stabilize actin filament, its interaction with TTC9A may play a role in cell shape and motility. In our previous results, we have found that progesterone-induced TTC9A expression was associated with increased cell motility and cell spreading. We speculate that TTC9A acts as a chaperone protein to facilitate the function of tropomyosins in stabilizing microfilament and it may play a role in cancer cell invasion and metastasis.
Cao
Shenglan
S
School of Biological Sciences, Nanyang Technological University, Singapore. ashleycsl@gmail.com
Ho
Gay Hui
GH
Lin
Valerie C L
VC
D000818:Animals; D001943:Breast Neoplasms / Q000378:metabolism* / Q000473:pathology; D019556:COS Cells; D002522:Chlorocebus aethiops; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D009361:Neoplasm Invasiveness; D009362:Neoplasm Metastasis; D009419:Nerve Tissue Proteins / Q000378:metabolism / Q000502:physiology*; D010455:Peptides / Q000737:chemistry; D011374:Progesterone / Q000378:metabolism; D017434:Protein Structure, Tertiary; D014335:Tropomyosin / Q000737:chemistry* / Q000378:metabolism / Q000502:physiology*; D020798:Two-Hybrid System Techniques
D009419:Nerve Tissue Proteins; D010455:Peptides; C498783:TPM3 protein, human; C511653:TTC9 protein, human; D014335:Tropomyosin; D011374:Progesterone