Retinoblastoma
(Page Author:  Greg Hibbets)
 
Retinoblastoma is a tumor that arises in the retina and accounts for about three percent of cancer in children below the age of fifteen years. (Donaldson, et. al. 1993) Retinoblastoma occurs most frequently in children and has an annual incidence of approximately four children per million. Ninety five percent of cases are diagnosed before the age of five. (Shields, et. al. 1991)Retinoblastoma may be manifested unilaterally(one eye), or bilaterally(both eyes). There are two classes of retinoblastoma. The first class is germline retinoblastoma. The other class of retinoblastoma is the sporadic form of the disease.Most unilateral disease is sporadic whereas all bilateral disease cases are germline.

Phenotypically, the majority of children with retinoblastoma have a white pupil instead of the normal black pupil. Other signs of retinoblastoma include red eye, poor vision, and inflammation of tissue surrounding the eye.

 The gene for retinoblastoma lies on chromosome 13 within the q14 band (long arm) and usually is a result of a deletion within this band. Results vary with mutations in the Rb gene. This occurs when the mutated allele (Rb-) is inherited from one parent and a normal allele (Rb+) from the other parent. Tumors arise when a spontaneous mutation occurs in the normal allele rendering both copies of the gene nonfunctional.  This form occurs when two normal alleles are inherited and both alleles spontaneously mutate. As could be expected, germline forms are much more common because the chances of two autosomal mutations is much less likely. (Lodish, et. al. 1999)

   The retinoblastoma gene, denoted Rb, is a gene that codes for the Rb protein, which functions as a tumor suppressor. Normally, Rb protein is bound to the transcription factor E2F keeping it in the inactive state. When the cell is in mid G1 of the cell cycle, induced expression for Cyclin D, Cdk4, Cdk 6, and the transcription factor E2F occurs in order to prepare the cell for mitosis. The cyclin/Cdk complexes initiate the phosphorylation of Rb, which releases the protein from E2F. Phosphorylation occurs at the amino acid serine and the location of such phosphorylation sites is highlighted in RED in the amino acid sequence.(Swiss Prot)  Subsequently, E2F is converted to the active form.E2F is then able to stimulate its own synthesis and that of Cdk2 and cyclin E. These particular Cdk/Cyclin complexes further stimulate the phosphorylation of Rb allowing more E2F activity. After mitosis, the level of Cdk/Cyclin complexes falls, leading to the dephosphorylation of Rb. As the protein is dephosphoylated, it again combines with E2F rendering it inactive. When there is a defective Rb protein, E2F is unregulating. Consequently, cell division becomes continuous resulting in the growth of a tumor.

   The Rb gene consists of a total of 180,388 base pairs which includes 23 exons. However, upon processing, the messenger RNA consists of only 4740 base pairs. Once synthesized by ribosomes, the Rb protein has 928 amino acids with a molecular weight of 106159 Daltons. The start codon (RED) starts 343 base pairs into the mRNA and the stop codon (BLUE)appears at base number 3127. Another interesting feature of this gene is the repeated regions found in the code. One of these regions is between base pairs 123912-125501.(OMIM)Additionally, other mammals also have Rb1, even though it may be found on other chromosomes.

Retinoblastoma mRNA
BASE COUNT     1508 a    887 c    862 g   1483 t
ORIGIN


        1 ttccggtttt tctcagggga cgttgaaatt atttttgtaa cgggagtcgg gagaggacgg
       61 ggcgtgcccc gcgtgcgcgc gcgtcgtcct ccccggcgct cctccacagc tcgctggctc
      121 ccgccgcgga aaggcgtcat gccgcccaaa accccccgaa aaacggccgc caccgccgcc
      181 gctgccgccg cggaaccccc ggcaccgccg ccgccgcccc ctcctgagga ggacccagag
      241 caggacagcg gcccggagga cctgcctctc gtcaggcttg agtttgaaga aacagaagaa
      301 cctgatttta ctgcattatg tcagaaatta aagataccag atcATGtcag agagagagct
      361 tggttaactt gggagaaagt ttcatctgtg gatggagtat tgggaggtta tattcaaaag
      421 aaaaaggaac tgtggggaat ctgtatcttt attgcacgag ttgacctaga tgagatgtcg
      481 ttcactttac tgagctacag aaaaacatac gaaatcagtg tccataaatt ctttaactta
      541 ctaaaagaaa ttgataccag taccaaagtt gataatgcta tgtcaagact gttgaagaag
      601 tatgatgtat tgtttgcact cttcagcaaa ttggaaagga catgtgaact tatatatttg
      661 acacaaccca gcagttcgat atctactgaa ataaattctg cattggtgct aaaagtttct
      721 tggatcacat ttttattagc taaaggggaa gtattacaaa tggaagatga tctggtgatt
      781 tcatttcagt taatgctatg tgtccttgac tattttatta aactctcacc tcccatgttg
      841 ctcaaagaac catataaaac agctgttata cccattaatg gttcacctcg aacacccagg
      901 cgaggtcaga acaggagtgc acggatagca aaacaactag aaaatgatac aagaattatt
      961 gaagttctct gtaaagaaca tgaatgtaat atagatgagg tgaaaaatgt ttatttcaaa
     1021 aattttatac cttttatgaa ttctcttgga cttgtaacat ctaatggact tccagaggtt
     1081 gaaaatcttt ctaaacgata cgaagaaatt tatcttaaaa ataaagatct agatcgaaga
     1141 ttatttttgg atcatgataa aactcttcag actgattcta tagacagttt tgaaacacag
     1201 agaacaccac gaaaaagtaa ccttgatgaa gaggtgaata taattcctcc acacactcca
     1261 gttaggactg ttatgaacac tatccaacaa ttaatgatga ttttaaattc tgcaagtgat
     1321 caaccttcag aaaatctgat ttcctatttt aacaactgca cagtgaatcc aaaagaaagt
     1381 atactgaaaa gagtgaagga tataggatac atctttaaag agaaatttgc taaagctgtg
     1441 ggacagggtt gtgtcgaaat tggatcacag cgatacaaac ttggagttcg cttgtattac
     1501 cgagtaatgg aatccatgct taaatcagaa gaagaacgat tatccattca aaattttagc
     1561 aaacttctga atgacaacat ttttcatatg tctttattgg cgtgcgctct tgaggttgta
     1621 atggccacat atagcagaag tacatctcag aatcttgatt ctggaacaga tttgtctttc
     1681 ccatggattc tgaatgtgct taatttaaaa gcctttgatt tttacaaagt gatcgaaagt
     1741 tttatcaaag cagaaggcaa cttgacaaga gaaatgataa aacatttaga acgatgtgaa
     1801 catcgaatca tggaatccct tgcatggctc tcagattcac ctttatttga tcttattaaa
     1861 caatcaaagg accgagaagg accaactgat caccttgaat ctgcttgtcc tcttaatctt
     1921 cctctccaga ataatcacac tgcagcagat atgtatcttt ctcctgtaag atctccaaag
     1981 aaaaaaggtt caactacgcg tgtaaattct actgcaaatg cagagacaca agcaacctca
     2041 gccttccaga cccagaagcc attgaaatct acctctcttt cactgtttta taaaaaagtg
     2101 tatcggctag cctatctccg gctaaataca ctttgtgaac gccttctgtc tgagcaccca
     2161 gaattagaac atatcatctg gacccttttc cagcacaccc tgcagaatga gtatgaactc
     2221 atgagagaca ggcatttgga ccaaattatg atgtgttcca tgtatggcat atgcaaagtg
     2281 aagaatatag accttaaatt caaaatcatt gtaacagcat acaaggatct tcctcatgct
     2341 gttcaggaga cattcaaacg tgttttgatc aaagaagagg agtatgattc tattatagta
     2401 ttctataact cggtcttcat gcagagactg aaaacaaata ttttgcagta tgcttccacc
     2461 aggcccccta ccttgtcacc aatacctcac attcctcgaa gcccttacaa gtttcctagt
     2521 tcacccttac ggattcctgg agggaacatc tatatttcac ccctgaagag tccatataaa
     2581 atttcagaag gtctgccaac accaacaaaa atgactccaa gatcaagaat cttagtatca
     2641 attggtgaat cattcgggac ttctgagaag ttccagaaaa taaatcagat ggtatgtaac
     2701 agcgaccgtg tgctcaaaag aagtgctgaa ggaagcaacc ctcctaaacc actgaaaaaa
     2761 ctacgctttg atattgaagg atcagatgaa gcagatggaa gtaaacatct cccaggagag
     2821 tccaaatttc agcagaaact ggcagaaatg acttctactc gaacacgaat gcaaaagcag
     2881 aaaatgaatg atagcatgga tacctcaaac aaggaagaga aatgaggatc tcaggacctt
     2941 ggtggacact gtgtacacct ctggattcat tgtctctcac agatgtgact gtataacttt
     3001 cccaggttct gtttatggcc acatttaata tcttcagctc tttttgtgga tataaaatgt
     3061 gcagatgcaa ttgtttgggt gagtcctaag ccacttgaaa tgttagtcat tgttatttat
     3121 acaagatTGA aaatcttgtg taaatcctgc catttaaaaa gttgtagcag attgtttcct
     3181 cttccaaagt aaaattgctg tgctttatgg atagtaagaa tggccctaga gtgggagtcc
     3241 tgataaccca ggcctgtctg actactttgc cttcttttgt agcatatagg tgatgtttgc
     3301 tcttgttttt attaatttat atgtatattt ttttaattta acatgaacac ccttagaaaa
     3361 tgtgtcctat ctatcttcca aatgcaattt gattgactgc ccattcacca aaattatcct
     3421 gaactcttct gcaaaaatgg atattattag aaattagaaa aaaattacta attttacaca
     3481 ttagatttta ttttactatt ggaatctgat atactgtgtg cttgttttat aaaattttgc
     3541 ttttaattaa ataaaagctg gaagcaaagt ataaccatat gatactatca tactactgaa
     3601 acagatttca tacctcagaa tgtaaaagaa cttactgatt attttcttca tccaacttat
     3661 gtttttaaat gaggattatt gatagtactc ttggttttta taccattcag atcactgaat
     3721 ttataaagta cccatctagt acttgaaaaa gtaaagtgtt ctgccagatc ttaggtatag
     3781 aggaccctaa cacagtatat cccaagtgca ctttctaatg tttctgggtc ctgaagaatt
     3841 aagatacaaa ttaattttac tccataaaca gactgttaat tataggagcc ttaatttttt
     3901 tttcatagag atttgtctaa ttgcatctca aaattattct gccctcctta atttgggaag
     3961 gtttgtgttt tctctggaat ggtacatgtc ttccatgtat cttttgaact ggcaattgtc
     4021 tatttatctt ttattttttt aagtcagtat ggtctaacac tggcatgttc aaagccacat
     4081 tatttctagt ccaaaattac aagtaatcaa gggtcattat gggttaggca ttaatgtttc
     4141 tatctgattt tgtgcaaaag cttcaaatta aaacagctgc attagaaaaa gaggcgcttc
     4201 tcccctcccc tacacctaaa ggtgtattta aactatcttg tgtgattaac ttatttagag
     4261 atgctgtaac ttaaaatagg ggatatttaa ggtagcttca gctagctttt aggaaaatca
     4321 ctttgtctaa ctcagaatta tttttaaaaa gaaatctggt cttgttagaa aacaaaattt
     4381 tattttgtgc tcatttaagt ttcaaactta ctattttgac agttattttg ataacaatga
     4441 cactagaaaa cttgactcca tttcatcatt gtttctgcat gaatatcata caaatcagtt
     4501 agtttttagg tcaagggctt actatttctg ggtcttttgc tactaagttc acattagaat
     4561 tagtgccaga attttaggaa cttcagagat cgtgtattga gatttcttaa ataatgcttc
     4621 agatattatt gctttattgc ttttttgtat tggttaaaac tgtacattta aaattgctat
     4681 gttactattt tctacaatta atagtttgtc tattttaaaa taaattagtt gttaagagtc

 



 
Retinoblastoma protein:  RR1.  Amino acid Sequence

        10         20         30         40         50         60 
         |          |          |          |          |          | 
MPPKTPRKTA ATAAAAAAEP PAPPPPPPPE EDPEQDSGPE DLPLVRLEFE ETEEPDFTAL

        70         80         90        100        110        120 
         |          |          |          |          |          | 
CQKLKIPDHV RERAWLTWEK VSSVDGVLGG YIQKKKELWG ICIFIAAVDL DEMSFTFTEL

       130        140        150        160        170        180 
         |          |          |          |          |          | 
QKNIEISVHK FFNLLKEIDT STKVDNAMSR LLKKYDVLFA LFSKLERTCE LIYLTQPSSS

       190        200        210        220        230        240 
         |          |          |          |          |          | 
ISTEINSALV LKVSWITFLL AKGEVLQMED DLVISFQLML CVLDYFIKLS PPMLLKEPYK

       250        260        270        280        290        300 
         |          |          |          |          |          | 
TAVIPINGSP RSRRGQNRS ARIAKQLEND TRIIEVLCKE HECNIDEVKN VYFKNFIPFM

       310        320        330        340        350        360 
         |          |          |          |          |          | 
NSLGLVTSNG LPEVENLSKR YEEIYLKNKD LDARLFLDHD KTLQTDSIDS FETQRTPRKS

       370        380        390        400        410        420 
         |          |          |          |          |          | 
NLDEEVNVIP PHTPVRTVMN TIQQLMMILN SASDQPSENL ISYFNNCTVN PKESILKRVK

       430        440        450        460        470        480 
         |          |          |          |          |          | 
DIGYIFKEKF AKAVGQGCVE IGSQRYKLGV RLYYRVMESM LKSEEERLSI QNFSKLLNDN

       490        500        510        520        530        540 
         |          |          |          |          |          | 
IFHMSLLACA LEVVMATYSR STSQNLDSGT DLSFPWILNV LNLKAFDFYK VIESFIKAEG

       550        560        570        580        590        600 
         |          |          |          |          |          | 
NLTREMIKHL ERCEHRIMES LAWLSDSPLF DLIKQSKDRE GPTDHLESAC PLNLPLQNNH

       610        620        630        640        650        660 
         |          |          |          |          |          | 
TAADMYLSPV RSPKKKGSTT RVNSTANAET QATSAFQTQK PLKSTSLSLF YKKVYRLAYL

       670        680        690        700        710        720 
         |          |          |          |          |          | 
RLNTLCERLL SEHPELEHII WTLFQHTLQN EYELMRDRHL DQIMMCSMYG ICKVKNIDLK

       730        740        750        760        770        780 
         |          |          |          |          |          | 
FKIIVTAYKD LPHAVQETFK RVLIKEEEYD SIIVFYNSVF MQRLKTNILQ YASTRPPTLS

       790        800        810        820        830        840 
         |          |          |          |          |          | 
PIPHIPRSPY KFPSSPLRIP GGNIYISPLK SPYKISEGLP TPTKMTPRSR ILVSIGESFG

       850        860        870        880        890        900 
         |          |          |          |          |          | 
TSEKFQKINQ MVCNSDRVLK RSAEGSNPPK PLKKLRFDIE GSDEADGSKH LPGESKFQQK

       910        920      928 
         |          |        |
LAEMTSTRTR MQKQKMNDSM DTSNKEEK

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