CELLS: THE LIVING UNITS-2

RIBOSOMES Universal cell organelles - Ribosomes. Cell organelles concerned with protein synthesis - Ribosomes (hence called protein factories). Ribosomes were discovered by Palade (Hence called Palade granules) in animal cell called them as microsomes. Claude called these structures as ribosomes. Membrane less cell organelles - Ribosomes. Ribosomes are the smallest cell organelles (230Å in diameter). Eukaryotic ribosomes are - 80S & 70S ribosomes. 80S ribosomes are found in cytoplasm. 70S ribosomes are found in Mitochondria. Ribosomes have two unequal sub units. 80S Ribosomes -> 40 S and 60 S subunits. 70S Ribosomes -> 30 S and 50 S subunits. Sub units of ribosomes are composed of - proteins & RNA (Hence called Ribonucleo protein particles). In eukaryotes, ribosomes are synthesized in Nucleolus (Ribosome factory). 60S sub units are attached to ER by means of proteins called - Ribophorins. 60S sub unit consists of - 40 proteins, 3 bits of rRNA (28S, 5.8S, & 5S). 40S sub units consists of - 30 proteins & a bit of (18S) rRNA. Chemical composition of 80S ribosomes - 40-60% RNA + 60-40% proteins. Sub units of ribosomes are combined by - Mg²+. A group (6 -8) of ribosomes attached to the same mRNA is called- Polyribosomes/Polysomes (or) ergosomes. Ribosomes are concerned with translation phase of protein synthesis. Ribosomes can be isolated by - cell Fractionatlon (homogenization & ultra centrifugation). (Rich, 1963). Centres of protein synthesis; site of protein synthesis; site of translation - Ribosomes. Enzyme for peptide bond formation is - Peptidyl transferase. Peptidyl transferase is located in - Larger subunit of ribosome (60 S or 50 S). Distinct sites in the larger sub unit are A - site (aminoacyl tRNA site). P - site (peptidyl tRNA site). Protein factories or work benches of proteins Ribosomes. MICRO ORGANELLES Lysosomes, Peroxisomes, Glyoxysomes, Sphaerosomes are together called Micro-organelles or Microbodies. All the micro organelles are single membrane bound structures. Lysosomes originate from Golgi complex, other micro-organelles develop from ER.

LYSOSOMES Lysosomes were discovered by Christian de Duve (1955) in rat liver cells. Lysosomes are called enzyme packets since they store several (50) types of hydrolysing enzymes to digest (autolysis) almost every type of organic matter except cellulose. Lysosomes are abundant in digestive glands. The most important enzyme in lysosome is acid phosphatase. Function of lysosome - digestion. Digestion of food, foreign bodies, etc., by lysosomes is called - heterophagy. Digestion of the own cell components by lysosomes is called - autophagy/autolysis. Lysosomes are called suicidal bags or atom bombs or disposal units or scavenger of cell. Lysosomes carry on Intracellular and extra cellular digestion. Lysosomes are polymorphic having different types. Lysosomes having only digestive enzymes - primary lysosomes. Lysosomes enclosing food vacuoles, foreign bodies or cell organelles are called - secondary lysosomes. Lysosomes with undigested matter are called Residual bodies/Tertiary Lysosomes. Size of lysosomes 0.2 -0.8m in diameter. PEROXISOMES Peroxisomes are microbodies which are also known as small lysosome which are mostly found in liver cells and help in detoxification of toxic substances inside the liver like – H2O2, alcohol etc. MICROTUBULES & MICRO FILAMENTS Micro tubules were discovered by - Ledbetter & Porter. Micro tubules are made up of a protein called Tubulin. Structures organized by microtubules - Spindle fibres, flagella, cilia, centrioles, cytoskeleton. Microtubules as spindle fibres determine the plane of new cell wall formation. Microtubules, intermediate filaments and microfilaments together form a new work called Cytoskeleton. CENTROSOME AND CENTRIOLES Centrosome is an organelle usually containing two cylindrical structures called centrioles. They are surrounded by amorphous peri centriolar materials. Both the centrioles in a centrosome lie perpendicular to each other in which each has an organisation like the cart wheel. They are mad up of nine evenly spaced peripheral fibrils, of tabulin protein. FUNCTION Centrioles form the basal body of cilia or flagella and spindle fibres that give rise to spindle apparatus during cell division in animal cell.

CILIA AND FLAGELLA Cilia and flagella are hair like out growths of cell membrane. Cilia are small structure with work like oars causing the movement of either the cell or the surrounding fluid. Flagella or cilia have three parts : (i) Root (ii) Basal plate (iii) Shaft NUCLEUS Dynamic centre of the cells is - Nucleus. Nucleus was discovered by- Robert Brown (1831) in orchid cells. Organisms with a true nucleus are called - Eukaryotes. Cells are usually uninucleate (with a single nucleus). Cell without nucleus - Enucleate cell. The study of nucleus is Karyology. Fusion of nucleus karyogamy and division is karyokinesis. Cellular activities and morphology are regulated by - nucleus. The vehicle of heredity is - nucleus. In higher organisms somatic cells have diploid nucleus (definitive nucleus) and sex cells have haploid nucleus Interphase nucleus is convenient for the study of its structure. Nucleus consists of i) Nuclear envelope, ii) Nucleoplasm iii) Nucleolus & iv) Chromatin reticulum. Nuclear envelope (karyotheca) is a double membraned envelope. The fluid filled space between the nuclear membranes perinuclear space (100-3000A°). Nuclear envelope in eukaryotes disappears during prophase, reappears during telophase. Nuclear envelope originates from ER (annulated RER). Transport of materials between cytoplasm and nucleoplasm occurs through nuclear pores. Nuclear pore +annulus (Protein cylinder) are together called nuclear pore complex. Intranuclear part of protoplasm is called - nucleoplasm. Nucleoplasm is also called nuclear sap, karyolymph, Karyoplasm, etc. Spherical membrane less structure found in the nucleus is called nucleolus. Nucleolus was discovered by Fontana.(1781) Nucleolus consists of DNA, RNA and proteins. Nucleolus is concerned with biogenesis of Ribosomes (Hence called Ribosome factory). Secondary constriction associated with nucleolus formation is called nucleolar organizer region. DNA and Histones together form chromatin. Chromatin fibres together appear as a net work called - Chromatin reticulum. Nucleus consists of a constant number of Chromatin fibres. Chromatin fibres condense to form chromosomes during prophase of cell division. The tightly coiled and deeply stained chromatin of interphase nucleus is called heterochromatin. The less stained and uncoiled part of chromatin is called Euchromatin. Euchromatin has active genes and heterochromatin has inactive genes. The terms Euchromatin & Heterochromatin were coined by Heitz. Lumps of heterochromatin are called Chromocentres, Karyosomes, pseudonucleoli.

CHROMOSOMES Rod shaped, deeply stained nucleoprotein bodies found in a dividing cell - Chromosomes. Chromosomes were discovered by Hofmeister in pollen mother cells of Tradescantia. The term ' Chromosome' was coined by Waldeyer. Chromosomes are visible during Mitosis and Meiosis. Best phase to observe chromosome morphology - Metaphase. Number of chromosomes is constant for each species. Best stage to count the chromosome number Diakinesis. Shape of the chromosome can be observed in Anaphase. STRUCTURE OF THE CHROMOSOME Longitudinal halves of a metaphase chromosome are called - Chromatids. Chromatids of the same chromosome (sister chromatids) remain attached at a point called primary constriction or centromere. Colourless constriction in the chromosome at which spindle fibres attach with the chromosome is called - primary constriction/centromere/Kinetochore/Kinomere. Centromere is useful for orientation of chromosomes at equitorial plane of the cell and their movement to opposite poles. Protein discs present on either side of centromere are also called - Kinetochores. Chromatic portions on either side of centromere are called - arms. Chromosome morphology is distinct with regard to relative lengths of arms, and position of centromere. Chromosome classification is based on the number of centromeres. Acentric - without centromere Monocentric - with one centromere Dicentric - with two centromeres Polycentric - with many centromeres Holocentric - with diffuse centromere Neocentric - with newly acquired centromere Usually chromosomes are monocentric. Chromosomes with genetically identical arms are isochromosomes (Darlington 1940). Constriction in the chromosome at which spindle fibres do not attach - Secondary constriction. Secondary constriction involved in nucleolus formation - nucleolar organizer region. Nucleolar organizer region contains - r RNA genes. Small spherical portion of chromosome distal to secondary constriction is called - Satellite/Trebent. Chromosome with satellite - SAT - Chromosome. The ends of chromosomes are called - Telomeres. Telomeres exhibit polarity and provide stability for chromosomes. Folded fibre model of chromosome was proposed by - Du Praw. De Robertis (1956) and Ris (1957) proved that pellicle and matrix are absent in the chromosome. Each chromatid has single Chromonema (mononemic).

CHEMICAL COMPOSITION OF CHROMOSOME Chromosomes contain DNA, Histones, RNA and non histone proteins. Histones are basic proteins. 5 types (H1, H1A, H2B, H3 and H4). Repeated units of chromatin - Nucleosomes. The term 'nucleosome' was coined by Wood-cock(1973). Nucleosome model was proposed by R.Kornberg. Chromosomes concerned with determination of body characters - Autosomes. Chromosomes concerned with sex of the organism are called Sex chromosomes/allosomes/heterosomes Eg. X, Y Chromosomes Sex chromosomes were discovered by C.E.Mc Clung. X - linked inheritance is called Sex linked inheritance. Characteristic chromosome complement of a species observed at metaphase of somatic cell is called Karyotype. Diagrammatic representation of Karyotype is called - Idiogram. Chromosomes of abnormal size are called giant chromosomes. FUNCTIONS Chromosomes are the Physical carriers of heredity or Vehicles of heredity. Chromosomal theory of inheritance was proposed by - W.S. Sutton (1902) and Theodore Boveri. Hereditory units are genes. Term 'gene' was coined by Johannsen. Gene theory (that genes are arranged in a linear order in the chromosome) was proposed by T.H.Morgan. Specific position of a gene in the chromosome is called - Locus. Genes which occupy the same locus in homologous chromosomes are called - alleles. Alternative forms of the same gene are called alleles. Genes found attached together in the same chromosome are called - Linkage group. Number of linkage groups in the organism is equal to the haploid number of chromosomes. All genes found in a haploid set of chromosomes are together called - genome. Genetic constitution of an organism is called - genotype. NUCLEIC ACIDS Nucleic acids are of two types - DNA & RNA (names were coined on the basis of sugar). Nucleic acids were discovered by - Friedrick Miescher from the nuclei of pus cells. Miescher called them nucleins and Altmann called them nucleic acids. Role of nucleins in heredity was suggested by Hertwig. Nucleic acids are unbranced, linear polymers of nucleotides. Structural units or building blocks of nucleic acids - nucleotides. Elements found in nucleic acids -C, H, 0 N, & P.

Nucleotides are made up of a pentose sugar + N2, base + Phosphate radicals. Nucleoside consists of a pentose sugar and N2 base. The bond between sugar and N2 base is called glycosidic bond. The bond between sugar and phosphate phosphodiester bond. Nitrogen bases are of two types - purines & pyrimidines. Dicyclic N2, bases with two carbon - nitrogen rings are called purines (Adenine & Guanine). Monocylic N2 Bases with a single carbon - nitrogen ring are called - pyrimidines (Thymine, Cytosine & Uracil). In the polynucleotide chain, nucleosides are joined by Phosphodiester bonds. DNA DNA means Deoxyribonucleic acid. Biochemical with highest molecular weight and utmost importance - DNA. DNA is the genetic material. It is proved by transformation experiments on Pneumococcus conducted by Avery et al and by experiments on T2 phage by Hershey and Chase. DNA double helix model was proposed by J.D.Watson & F.H.C. Crick (1953). Scientists awarded Nobel Prize in 1962 for DNA structural model are Watson, Crick and Wilkins. Sugar in DNA is deoxyribose sugar. N2 bases in DNA are - Adenine, & Guanine (purines) Thymine and Cytosine (Pyrimidines). DNA is a right handed double helix. The two polynucleotide strands are coiled around a common axis in clockwise direction. The two polynucleotide strands of DNA are quite complementary and antiparallel. The polynucleotide strands are held together by hydrogen bonds. Purines & pyrimidines exist in 1:1 ratio which is due to complete complimentarity between polynucleotide strands. Length of DNA molecule is variable and diameter is constant (20 Å). Length of one coil of DNA is 34 Å. No. of nucleotides in one coil of DNA is 10 pairs or 20 nucleotides. Distance between successive nucleotide pairs is 3.4 Å and angle between them is 36°. DNA self replication takes place by Semiconservative method. Semiconservative method of DNA self replication was proposed by Watson & Crick and proved experimentally by Messelson and Stahl using N15 isotope in E.coli. Enzyme useful for DNA self replication is DNA polymerase. Synthesis of m RNA from DNA template is the heterocatalytic function of DNA. Proteins associated with DNA of eukaryotes are histones. Single stranded DNA is found in φ × 174 phage. Artificial DNA synthesis was carried by Arthur Kornberg. Artificial gene synthesis was carried by H.G. Khorana. Circular naked DNA is found in Mitochondria.

RNA RNA means Ribonucleic acid. RNA is a single stranded polymer of ribonucleotides. Double stranded RNA is found in certain viruses - Reo virus, Wound tumour virus. RNA is nongenetic material. But in viruses it is the genetic material eg. TMV. N2 bases in RNA are - Adenine guanine, uracil and cytosine. In RNA Uracil replaces Thymine. Chargaff's rules are not applicable to RNA since it is single stranded. RNA is synthesized from DNA. The function of nongenetic RNA is protein synthesis. Non genetic RNA is of 3 types - m RNA, r RNA & tRNA. Largest type of RNA is ribosomal RNA (rRNA)and the smallest one is transfer RNA (t RNA). RNAs are produced from template DNA as complementary strands. Enzyme catalysing RNA synthesis is RNA Polymerase. mRNA Formation of m RNAfrorn DNA template is called Transcription. Direct template for protein synthesis is m RNA. m RNA was discovered by Jacob & Monod. Genetic information for protein synthesis is found in m RNA in the form of triplet codes or codons. Codon is a sequence of 3 successive nucleotides. m RNA constitutes 2-5% of total cellular RNA. m RNA is unstable. tRNA tRNA is also called soluble RNA, adopter RNA or interpreter of genetic code. tRNA consists of about 75-85 nucleotides (smallest type of RNA). tRNA is stable and constitutes about 15% of total RNA. tRNA is useful to bring suitable aminoacids into ribosomes during protein synthesis. Clover leaf model for RNA was proposed by R. Holley. tRNA with attached amino acid is called charged t RNA/aminoacyl t RNA. tRNA recognises the codon in m RNA by its anticodon. Three dimentional (3D) L – shaped model for t RNA was proposed by S.H. Kim, et al. rRNA Ribosomal RNA (r RNA) is a component part of ribosomal sub units. rRNA is the largest type of RNA and has several thousands of nucleotides. rRNA constitutes about 80% of total RNA. rRNA is in the form of an irregular double helix. Enzymes which destroy nucleic acids are nucleases (DNase, RNase).

CELL CYCLE
Reproduction at cellular level is called cell division. Cell division was first studied by Strasburger in plants. W. Fleming (1882) in animal cells and Prevost and Dumas (1824) in frog egg. 2. All organisms start their life from a single Primordial cell called Zygote. 3. In Multicellular Animals growth and Development take place due to cell division. Mitogens are the agents which stimulate cell division. Mitotic poisons are inhibitors of cell division. Azides and cyanides inhibit prophase,Colchicine checks spindle formation ,Chalones inhibit cell division in vivo and in vitro both ,Ribo nuclease blocks prophase, Heat shock prevent cell division and Mustard gas agglutinate all chromosomes. 4. Cell division is divided mainly into two phases (a) Karyokinesis (b) cytokinesis 5. Division of nucleus is Karyokinesis Division of cytoplasm is cytokinesis. Animal cytokinesis is centripetal. 6. Karyokinesis is of two types : (1) Mitosis (2) Meiosis MITOSIS OSIMitosis is called Equational Division because it results in the formation of two identical daughter Nuclei It is also called Indirect Nuclear division because of the formation of spindle and visible changes in chromosomes. It is also called Somatic Nuclear division because it occurs in the Nucleus of Somatic cells. The term Mitosis was used by Fleming in 1882 Mitosis is divided into two phases (1) Inter phase (2) Division phase (M - phase) INTERPHASE The interval between two division phases is called Interphase It is the longest phase in Mitotic cycle. It is also called as Resting phase Metabolically active stage of Mitosis is Interphase. Period of Intense biosynthetic activity is interphase Interphase is divided into 3 phases G1 phase, S phase, G2 phase G1phase is called First growth phase, gap period, Presynthetic Period ) S phase is synthetic phase, Replication of DNA (Chromosomes) occurs during S phase. G2 phase is Second growth phase or Post synthetic Period. Metabolic activities like growth of cytoplasm and its constituents occur during this phase. Interphase takes one or two days for its completion. CHANGES IN INTERPHASE Nucleus is large and distinct Nucleus has got nuclear membrane Chromatin network is not distinguished into chromosomes. Nucleolus is also prominently visible . 1.

DIVISION PHASE (M - PHASE) Time taken for completion of a division is called division phase Time interval for completing a division as well as for its preliminary preparation is called cell cycle. Cell cycle includes Interphase and a Division phase. For convenience of study division phase is divided into 4 phases. 7. M- Phase:it is having following stages PROPHASE The chromatin network begins to coil and appear as longthread-like structures called as chromosomes. Each chromosome consists of two chromatids. They lie side by side and are join at a point called as centromere. The nucleolus gradually disappears from early prophase to late prophase The nuclear membrane also starts disappearing in the late prophase. METAPHASE It starts with the complete disappearance of Nuclear Membrane The chromatids become shorter and thicker due to dehydration and condensation Chromatids acquire a specific shape & size. The chromatids move to the centre of the cell with their centromeres this is called metakinesis Five Fibrils appear in the cell cytoplasm and are organised to form spindle fibres. Asters are formed. The centromere of each chromosome lies on the equator and is attached to the spindle fibres. The best stage of Mitosis for analysing the chromosomes is Metaphase ANAPHASE The centromere of each chromosome is divided into two . So each chromatid get its own centromere. Chromatid with its own centromere is regarded as a Daughter Chromosome. The spindle fibres get attached to the centromeres of their side The spindle fibres gradually shortens, by this each chromatid with its centromere is pulled apart towards its respective pole. At the poles, each chromatid now behaves as Independent chromosome so the chromosome number of daughter nuclei is maintained the same The centromeres of the chromosomes lie towards the poles of the spindle, where as the arms are directed towards the Equator, At the End of Anaphase, a constriction appears in the middle periphery of the cytoplasm. TELOPHASE Last phase of Karyokinesis is Telophase Changes occuring during Telophase are reverse to Changes occured during prophase. At each pole, the chromatids (now the Daughter chromosomes) become uncoiled , thin and invisible. Chromosomes are again reorganised into chromatin Network The Nucleolus and nuclear membrane reappears again Thus two daughter nuclei are formed at the two poles of a cell. The formed two daughter nuclei are qualitatively and quantitatively similar to the parent Nucleus. Finally the cell is simply Pinched off into two daughter cells by a constriction

8.

Meiosis: The cell division in which chromosome number is reduced / to half is called Meiosis. Meiosis was discovered by Strasburger. The Term Meiosis was used by Farmer and Moore Meiosis must take place in an organism to maintain the chromosome number constant. In animal cells Meiosis is termed as “Terminal or gametic meiosis” The cell which undergoes Meiosis is called a Meiocyte In meiosis one Replication of chromosome is followed by two divisions in the nucleus and cytoplasm

MEIOSIS – (FIRST NUCLEAR DIVISION) In this division, the chromosome number is reduced to half. New daughter cells produced are dissimilar to the parent cell. It is also called Reductional division or Heterotypic division

IT IS DIVIDED INTO THE FOLLOWING STAGES
Prophase I It takes long duration and differs from prophase of mitosis. It is divided into 5 subphases. 9. (a) Leptotene or Leptonema :The nucleus increases in size and volume by absorbing water The chromatin network shows condensation and forms a number of long thread like structures called as chromosomes. Each chromosome shows characteristic beaded appearance because of the presence of chromomeres along the entire length Nuclear membrane and Nucleolus are distinct. 10. (b) Zygotene or Zygonema :The two homologous chromosomes (one paternal and one maternal ) lie side by side. This is known as pairing or Synapsis of Homologous chromosomes. Each pair of chromosomes is known as Bivalent Further shortening of chromosomes is present due to the coiling and chromosomes become more distinct Nuclear membrane and nucleolus are present 11. (c) Pachytene or pachynema Each chromosome of a bivalent splits longitudinally into two sister chromatids by this Bivalent becomes a tetrad, The two chromatids one from each bivalent partially coil around each other of exchange their genetic material This is known as crossing over. The points of crossing over are known as chiasmata. Nucelolus and Nuclear membrane are present. 12. (d) Diplotene or Diplonema Homologous chromosomes which had paired during Zygotene now start separating from one another As the chromosomes are showing gradual condensation, so there is a tendency that chiasmata tend to slip out of the chromosomes This is known as Terminalisation of chiasmata. Chromosomes start seperating out but the separation is not complete Nuclear membrane and Nucleolus start degenerating. 13 . (E) Diakinesis The Bivalents condense further and get randomly distributed in the cytoplasm. The separation of paired chromosomes is almost complete Terminalisation of chiasmata is almost complete. Nuclear membrane and Nucleolus disappear 14. Metaphase I The bivalents come to lie in the Equatorial plane in such a way that one member faces one pole of the spindle apparatus, and the other member on the other pole. Thus two members of the pair lie on opposite side of the equator. Nuclear membrane and nucleolus completely disappear. Centromeres of the chromosomes are attached to spindle fibres

15. Anaphase - I Each chromosome, consisting of two chromatids, separate and move to opposite poles as they are pulled apart by the shortening of spindle fibre. The number of chromosomes moving towards the opposite poles is reduced to half. The chromosomes on the poles are genetically different from the parent chromosomes due to crossing over. 16. Telophase -I At the opposite poles, the chromosomes loose their identity and Nuclear membrane is formed around the nuclei Nucleolus also reappears Thus each nucleus formed has half the number of chromosomes as compared to the parent cell Meiosis –I results in the formation of two haploid nuclei from a diploid nucleus Meiosis – II is also called Homotypic Division or Equational division It is similar to mitosis and divided into following four stages. 17. Prophase II The chromosomes reappear Each chromosome has two chromatids Nucleus membrane and nucleolus disappear The chromosome become short by coiling and condensation. 18. Metaphase II The chromosomes get arranged on the equator of the spindle. Centromere divides into two The microtubules of the spindle are attached with the centromere of the chromosomes. 19. Anaphase II The centromere of each chromosome divides into two so that now each chromatid gets its centromere Each centromere gets attached to the spindle fiber of its side Shortening of spindle fibers occurs so that the chromatids are pulled apart towards their respective poles Nucleolus and Nuclearmembrane are absent. 20. Telophase II The chromatids (called Daughter chromosomes ) on the respecitve poles uncoil and form the chromatin network again. The Nuclear membrane and Nucleolus are formed Each Haploid nuclei are seen in each cell Finally cytoplasm is constricted into four parts to gives four daughter cells So after Meiosis - II 4 haploid daughter cells are formed. PROTEIN SYNTHESIS Protein synthesis take place with the help of DNA, RNA and ATP molecules. Protein synthesis takes place in two steps. (i) Transcription (ii) Translation (i) Transcription : Formation of RNA by the DNA is known as transcription. In this process two strands of DNA uncoil. In which one of them function like template for the synthesis of RNA strand. In the formation of new RNA strand N2 Base arranged according to the sequence of DNA template at the place of thymine, uracil attached at the RNA strand. In this manner new RNA form which comes in to cytoplasm and bring the coded information in the form of genetic code which are triplet i.e. (AAG, AUG, AUC, etc.) (ii) Translation : Formation of the chain of aminoacid on the ribosome with the help of mRNA , transfer RNA and ATP molecules is known as translation. Triplet code of mRNA which brings from the DNA functions like the code for amino acid.

Process of Translation :The m-RNA is attached to the groove of two sub unit of ribosome. On the basis of triplet code transfer RNA bring aminoacid with the help of ATP to the ribosome. On the ribosome amino acid attached one by one from initiation codon to terminal codon and form a polypeptide chain. Ultimately one or more than one polypeptide chain attain their 3-D structure and form protein molecules.

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