Molecular analysis of CIB4 gene and protein in Kermani sheep.

M.R Mohammadabadi,F Bordbar,A.H.D Jafari

doi:10.1590/1414-431x20176177

M.R Mohammadabadi, F Bordbar + Show 1 more

Open Access

https://doi.org/10.1590/1414-431x20176177

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Abstract

The human calcium- and integrin-binding protein (CIB) family is composed of CIB1, CIB2, CIB3, and CIB4 proteins and the CIB4 gene affects fertility. Kermani sheep is one of the most important breeds of Iranian sheep breeds. The aim of this study was to analyze for the first time molecular characteristics of the CIB4 gene and protein in Kermani sheep. Different tissues were collected from the Kermani sheep and real time PCR was performed. The PCR products were sequenced, comparative analyses of the nucleotide sequences were performed, a phylogenetic tree was constructed, and different characteristics of CIB4 proteins were predicted. Real time PCR results showed that the CIB4 gene is expressed only in testis of Kermani sheep. The cDNA nucleotide sequence was identical with small tail Han sheep, cattle, goat, camel, horse, dog, mouse and human, respectively 100, 99, 99, 98, 98, 96, 96, and 96%. Hence, it can be suggested that the CIB4 gene plays a role in male fertility. Based on the phylogenetic analysis, sheep CIB4 gene has a close relationship with goat and cattle first, and then with camel and whale. Although we demonstrated that CIB4 is a testis-specific gene, expressed only in the testis and it interacts with other proteins, the mechanisms by which CIB4 expression is regulated need to be elucidated.

Highlights

The human calcium- and integrin-binding protein (CIB) family is composed of CIB1, CIB2, CIB3, and CIB4 proteins and their molecular weight is approximately 22 kDa
Researchers demonstrated that a CIB1 gene knockout mouse displays no overt defect in platelet function [2], which suggests that CIB2, CIB3, and CIB4 members can compensate for CIB1 loss
All RNA extracted from brain, heart, lung, spleen, kidney, liver, ovary, and testis of the Kermani sheep used in the present study revealed two 18S and 28S bands (Figure 1)

Summary

Introduction

The human calcium- and integrin-binding protein (CIB) family is composed of CIB1, CIB2, CIB3, and CIB4 proteins and their molecular weight is approximately 22 kDa. It has been proven that mammalian CIB1 functionally affects hemostasis, DNA damage response, apoptosis, embryogenesis [1], endomitosis [7], and spermatogenesis [8]. Other names for this protein are KIP [9] and calmyrin [10]. It has newly been proven that the CIB2 gene is expressed in the brain of rat [11] and in skeletal muscle of mouse, where the produced protein can interact with the integrin a7B subunit [12]. It has been proven that the CIB2 gene is expressed in various sheep tissues, including liver, heart, kidney, brain, spleen, stomach, ovary, testis, and muscle [13]. It has been shown that the CIB4 gene expresses only in the testis [3]

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