Hsiao K C, Cheng C H, Fernandes I E, Detrich H W, DeVries A L
Department of Physiology and Biophysics, University of Illinois, Urbana 61801.
Proc Natl Acad Sci U S A. 1990 Dec;87(23):9265-9. doi: 10.1073/pnas.87.23.9265.
The antarctic fish Notothenia coriiceps neglecta synthesizes eight antifreeze glycopeptides (AFGP 1-8; Mr 2600-34,000) to avoid freezing in its ice-laden freezing habitat. We report here the sequence of one of its AFGP genes. The structural gene contains 46 tandemly repeated segments, each encoding one AFGP peptide plus a 3-amino acid spacer. Most of the repeats (44/46) code for peptides of AFGP 8; the remaining 2 code for peptides of AFGP 7. At least 2 of the 3 amino acids in the spacers could act as substrate for chymotrypsin-like proteases. The nucleotide sequence between the translation initiation codon (ATG) and the first AFGP-coding segment is G + T-rich and encodes a presumptive 37-residue signal peptide of unusual sequence. Primer extension establishes the transcription start site at nucleotide 43 upstream from ATG. CAAT and TATA boxes begin at nucleotides 53 and 49, respectively, upstream from the transcription start site. The polyadenylylation signal, AATAAA, is located approximately 240 nucleotides downstream from the termination codon. A mRNA (approximately 3 kilobases) was found that matches the size of this AFGP gene. Thus, this AFGP gene encodes a secreted, high-copy-number polyprotein that is processed posttranslationally to produce active AFGPs.
南极鱼类小头南极鱼(Notothenia coriiceps neglecta)合成了8种抗冻糖肽(AFGP 1 - 8;分子量2600 - 34,000),以避免在充满冰的寒冷栖息地中被冻住。我们在此报告其一种AFGP基因的序列。该结构基因包含46个串联重复片段,每个片段编码一个AFGP肽加上一个3个氨基酸的间隔区。大多数重复片段(44/46)编码AFGP 8的肽段;其余2个编码AFGP 7的肽段。间隔区的3个氨基酸中至少有2个可作为类胰凝乳蛋白酶的底物。翻译起始密码子(ATG)与第一个AFGP编码片段之间的核苷酸序列富含G + T,并编码一个序列异常的推定37个残基的信号肽。引物延伸确定转录起始位点在ATG上游43个核苷酸处。CAAT盒和TATA盒分别从转录起始位点上游53和49个核苷酸处开始。聚腺苷酸化信号AATAAA位于终止密码子下游约240个核苷酸处。发现了一种与该AFGP基因大小匹配的mRNA(约3千碱基)。因此,该AFGP基因编码一种分泌型、高拷贝数的多蛋白,该多蛋白在翻译后被加工以产生有活性的AFGPs。